Introduction: Focal Segmental Glomerulosclerosis (FSGS) is a heterogeneous glomerular disorder characterized by segmental sclerosis of one or more glomeruli and prominent proteinuria. Podocyte injury underlies the pathogenesis and compromises the renal filtration barrier. FSGS can be classified into primary, genetic, and secondary forms, with secondary causes including maladaptive, infection-related, and drug-induced mechanisms. These subtypes differ in etiology, clinical course, treatment response, and prognosis. Methods Whole exome sequencing was conducted on patients with biopsy-proven FSGS to identify the underlying molecular etiopathogenesis utilizing the Illumina NextSeq2000 system. Bioinformatic analyses were performed on the SOPHIA DDM platform. Sanger sequencing was conducted to validate the detected variant in the proband and to conduct segregation analysis in her family members. Results Among the 50 FSGS patients, no clinically relevant variants were identified by NGS in 25 individuals. Known pathogenic variants in SDCCAG8 , NPHS2 , and COQ8B were confirmed by Sanger sequencing. Three novel variants in MAPKBP1 , ARHGAP24 , and PODXL were identified and validated through familial segregation analysis. Variants in seven probands were confirmed by Sanger sequencing, while 12 novel VUS variants failed to segregate with disease in family studies. Conclusion This study highlights the value of genetic analysis in understanding its molecular basis. Integration of WES with detailed clinical evaluation allowed the identification of multiple variants, improving insight into FSGS pathogenesis. Limitations include the lack of functional validation for novel variants and the inability to perform segregation analyses in some families.

Amelogenesis imperfecta type 1G (AI1G) is a rare inherited condition caused by homozygous or compound heterozygous mutations in FAM20A. The disease was characterized by hypoplastic enamel on primary and secondary dentition, delayed or failed eruption of secondary dentition, pulp stones, gingival hyperplasia, and nephrocalcinosis. Loss-of-function mutation of FAM20A is associated with fibroblast growth factor 23 (FGF23)-mediated hypophosphataemia. Here, we present a case of AI1G presenting with hypophosphatemia. A 23-year-old male patient was referred to our clinic with fatigue, muscle weakness, and hypophosphatemia that was detected during an evaluation for incidentally discovered nephrocalcinosis. The patient's medical history included hypoplastic enamel, unerupted permanent teeth, and newly diagnosed hypertension. The image of the permanent teeth was shown in Figure 1. His parents were consanguineous. Family history was unremarkable. In examination, his length was 172 cm, and his weight was 65 kg. The patient had dental prostheses; otherwise, the physical examination findings were normal. Laboratory results showed creatinine, 1.2 mg/dL (normal, 0.7-1.4); phosphorus, 1.4 mg/dL (normal, 2.7-4.5); albumin, 4.8 g/dL (normal, 3.2-5.5); calcium, 9.2 mg/dL (normal, 8.5-10.5); PTH, 34 pg/mL (normal, 15-65); and ALP: 372 U/mL (normal, 90-220); 25 (OH) D level was normal, 24-hour urinary calcium, 26 mg; phosphorus, 950 mg; and creatinine, 1600 mg. Tubular maximum phosphate reabsorption per glomerular filtration rate (TmP/GFR) was calculated as 0.3 mmol/L (normal, 1-1.35) using Walton-Bijvoet nomogram. Laboratory revealed renal phosphate wasting. No glucosuria or proteinuria was detected. Bicarbonate level was normal; metabolic acidosis was not observed. Bone scintigraphy and abdominal and chest computed tomography scans were performed. No abnormalities were detected, except for bilateral nephrocalcinosis. Oral phosphate replacement and calcitriol therapy were initiated. The patient was monitored with serum calcium, phosphate, and PTH levels, renal function tests, and urinalysis, urinary ultrasonography. The doses of medication were adjusted according to plasma phosphate and PTH levels to prevent secondary hyperparathyroidism. Clinical exome sequencing identified a homozygous c.34_35del, p.(Leu12Alafs*67) frameshift mutation in exon 1 of the FAM20A gene. It was evaluated as a pathogenic mutation associated with AI1G (enamel-renal syndrome). After approximately 22 years of follow-up, the last laboratory results are summarized in Table 1. Hypophosphatemia has been reported rarely in AI cases. Although the mechanism of hypophosphatemia is not clearly understood, it has been suggested that it may arise through FGF23-mediated mechanisms. Additionally, measuring plasma phosphorus levels in patients with dental findings may provide further data regarding the presence and frequency of hypophosphatemia.Figure 1:Oral view of unerupted or partially erupted permanent teeth and dental prostheses Table 1:Laboratory results at the last follow-up

Summary We report that biallelic LRRK2 loss-of-function (LoF) causes a Mendelian form of interstitial lung disease characterized by alveolar epithelial cell dysfunction and lung fibrosis in two brothers with a homozygous nonsense variant. Integrated clinical, imaging, histopathological, and biomarker analyses showed absent LRRK2 protein, reduced Rab10 phosphorylation, impaired alveolar type 2 cell function, and disrupted surfactant homeostasis. Consistent with a recessive genetic disorder, heterozygous LoF carriers have not been reported to have a lung phenotype. Additional biallelic LRRK2 LoF cases were identified in ILD cohorts, linking LRRK2 LoF to lung disease, in contrast to heterozygous activating missense variants that cause Parkinson’s disease (PD).

Aim: This study aimed to investigate the functional effects of miR-15a-5p and miR-16-5p in AML cell lines (HL-60 and NB4), focusing on their regulation of key target genes.Material and Methods: We initially transfected miR-15a-5p and miR-16-5p mimics into AML cell lines to investigate their regulatory roles in fundamental cellular processes. After the transfection, the potential target genes of these miRNAs were identified through several in silico prediction tools, allowing us to focus on biologically relevant candidates. To evaluate how these miRNA–target gene axes may influence AML-related mechanisms, the expression levels of the selected genes were subsequently analyzed in mimic-transfected cells using quantitative expression assays.Results: Study results showed that transfection of miR-15a-5p and miR-16-5p miRNA mimics led to a marked suppression of HL-60 and NB4 AML cell proliferation, indicating that both miRNAs exert inhibitory effects on leukemic cell growth. Consistent with this observation, IGF1R expression was significantly downregulated at the mRNA level in AML cells transfected with either miRNA mimics. This coordinated reduction suggests that IGF1R may function as a critical downstream effector of these miRNAs.Conclusion: These findings suggest that miR-15a-5p and miR-16-5p may function as tumor suppressors in AML, with potential therapeutic implications. Additionally, the interactions between miR-15a-5p/miR-16-5p/IGF1R may serve as specific biomarkers for new therapeutic targets in AML. Overall, this study highlights miR-15a-5p and miR-16-5p as promising targets for the development of novel AML therapies.

Introduction: Hypouricemia is an often-overlooked condition. Isolated persistent hypouricemia is rare and may be associated with uncommon genetic disorders, such as familial renal hypouricemia or xanthinuria. Methods: Fifteen non-consanguineous adult patients were included in this single-center study. Secondary causes of hypouricemia, including malnutrition, SIADH, cirrhosis, uricosuric drug use, and full-blown Fanconi syndrome, were excluded. Patients were classified as hyperuricosuric or hypouricosuric based on urinary uric acid levels. Clinical or whole-exome sequencing was performed, and variant pathogenicity was assessed using in silico prediction tools. Results: Ten patients were female (66.7%), and four were hypouricosuric (26.7%). Three patients (20%) had also glucosuria without diabetes mellitus and full-blown Fanconi syndrome. Thirteen patients (86.7%) carried at least one rare variant (variant of unknown significance, likely pathogenic, or pathogenic) in genes associated with hypouricemia: 1 patient with homozygous SLC2A9, one with homozygous SLC22A12, five with heterozygous SLC22A12 (1 patient carried two variants; compound heterozygosity could not be confirmed), one with heterozygous CLCN5, one with homozygous XDH, two with homozygous MOCOS, one with heterozygous MOCS1, and one with two heterozygous SLC5A2 variants. This study reports, for the first time, the co-occurrence of familial renal glucosuria and familial renal hypouricemia, as well as the coexistence of xanthinuria type 2 and familial renal glucosuria in 2 patients. Finally, 1 female patient with hyperuricosuric hypouricemia carried a likely pathogenic FTL variant and a mitochondrial DNA variant of unknown significance, which may represent candidate genes and require confirmation in larger cohorts and functional studies. Conclusion: Our study expands the clinical and genetic spectrum of persistent hypouricemia. Genetic testing has a high diagnostic yield and should be considered in patients with unexplained persistent hypouricemia.

Purpose: This study aims to investigate the expression profiles of hsa_circ_0025244 and hsa_circ_0001546 in breast tissue samples and to propose potential regulatory axes for these circular RNAs. Materials and Methods: The expression levels of hsa_circ_0025244 and hsa_circ_0001546 were determined in 55 tumor tissue samples (30 Luminal B and 25 Luminal A) and 55 adjacent normal tissue samples using quantitative polymerase chain reaction. Subsequently, in silica databases and PubMed literature searches were employed to identify potential sponge microRNAs and target genes. Finally, potential regulatory axes were proposed for further investigation in breast cancer. Results: Both hsa_circ_0025244 and hsa_circ_0001546 were downregulated in total tumor tissues and Luminal B tumor tissues compared to normal tissues. However, only hsa_circ_0025244 was downregulated in Luminal A tumors compared to normal tissues. According to in silica tools and PubMed searches, the strongest target microRNA/gene association was determined to be hsa-miR-210-5p/BTG2 for hsa_circ_0025244 and hsa-miR-532-5p/ZFHX3 for hsa_circ_0001546. Conclusion: This study demonstrates that, hsa_circ_0025244 and hsa_circ_0001546, investigated for the first time in breast cancer, were found to exhibit decreased expression in breast cancer compared to normal tissue samples.

Aims: Larynx squamous cell carcinoma (LSCC) is the second most common head and neck malignancy. While let-7b-3p has been shown to have a role in cancer progression in malignancies, there is no research examining the association between LSCC and let-7b-3p. This study aimed to investigate the expression status of let-7b-3p and the potential roles of this microRNA (miRNA) in LSCC. Methods: Using quantitative real-time polymerase chain reaction (qRT-PCR), we examined the expression status of let-7b3p in 36 LSCC samples and the neighboring normal tissues. Then, the let-7b-3p miRNA mimic was transfected into Hep-2 cells via lipofectamine 2000 reagents. Cell viability was determined using the cell viability detection (CVDK-8) kit, and cell migration was evaluated with the scratch assay. To identify differentially expressed genes (DEGs) in larynx cancer GSE137308 and GSE130605 datasets were downloaded and reanalyzed using Gene Expression Omnibus (GEO2R) tool. Potential target genes of let-7b-3p were investigated in the miRNA target prediction and functional annotation database (miRDB). Shared genes between geo datasets and miRDB results were identified and the relationship between these genes and LSCC was investigated in the literature. Results: We demonstrated that the expression levels of let-7b-3p was significantly upregulated in LSCC tumor tissues in comparison to the corresponding normal tissues. Mimic let-7b-3p transfection enhanced Hep-2 cell proliferation and migration. In vitro and bioinformatics analysis showed that overexpression of let-7b-3p can enhance the larynx cancer cell proliferation and migration through MYBPC1. Conclusion: It was evaluated that let-7b-3p/MYBPC1 axis could potentially affect the LSCC process. Let-7b-3p has the potential to be a biomarker for LSCC, therefore, the let-7b-3p/ MYBPC1/LSCC relationship should be elucidated with new studies.

Objective: Acute Myeloid Leukemia (AML) is distinguished by the differentiation and overgrowth of blast cells. In the current study, we purposed to elucidate the effect of miR-7-5p on AML cellular processes and the expression level of potential target genes. Methods: miR-7-5p mimic was transfected into AML cells by lipofectamine-mediated method and verified by qRT-PCR. The miR-7-5p's effect on proliferation and apoptosis was investigated by WST-8 and Caspase-3 kit (respectively). miRDB, miRTarBase, Targetscan, miRWalk, https://ongene.bioinfo-minzhao.org/, and http://soft.bioinfo-minzhao.org/lgl/a databases were utilized for in silico identification of possible target genes of miR-7-5p. Relative gene expression of potential target genes was investigated via the qRT-PCR technique. Results: In the group that is transfected with miR-7-5p, proliferation significantly decreased and apoptosis increased as against the control group. BCL2, SKP2, OGT, KLF4 and EGFR gene expression levels, which were determined as a result of possible target gene analysis by in silico methods and literature search, were investigated in AML cell lines. While the SKP2, KLF4 and OGT expression levels were statistically decreased in the group of transfected with mimic miR-7-5p, no statistically significant change was detected in the expressions of BCL2 and EGFR genes. Conclusion: miR-7-5p may affect the cancer process in AML by targeting SKP2, KLF4, and OGT genes. It is very important to identify and validate the miR-7-5p target genes, which has the possibility to be a new biomarker in the early diagnosis and therapy of AML. Therefore, our data obtained at mRNA level should be confirmed by further studies.

Persistent hypouricemia (HU) (serum uric acid <2.5 mg/dL) is a rare condition. HU due to rare genetic causes, without full-blown Fanconi syndrome (FS) (characterized by glucosuria and hyperuricosuria alongside aminoaciduria, phosphaturia, and renal tubular acidosis), is typically associated with familial renal HU due to SLC12A22 and SLC2A9 variants, as well as xanthinuria linked to XDH and MOCOS variants. Nine patients were included. Malnutrition and full-blown FS were excluded. Clinical Exome Sequencing was performed, and pathogenicity of variants were evaluated with 1-SIFT, Polyphen2, and Mutation Taster prediction tools. 8 of 9 patients had at least one rare variant as a possible cause of HU. 5 patients had isolated HU, while 3 had both glucosuria and HU without other signs of FS. One patient with recurrent acute kidney injury had a homozygous variant in SLC2A9. Two patients with nephrolithiasis (NL) were diagnosed with xanthinuria type 1 and type 2. The patient with xanthinuria type 2 also had concurrent glucosuria due to additional heterozygous SLC5A2 variant. Homozygous and heterozygous SLC22A12 variants were detected in 2 patients with persistent HU. One patient with glucosuria and HU due to hyperuricosuria had no variants related to HU, but compound heterozygous SLC5A2 variants. The last case had a rare heterozygous FTL (Ferritin-light-chain) variant. This patient had hyperuricosuria. Transcriptomic and single-cell RNA databases showed that FTL has high expression in the proximal tubule, suggesting it may be a candidate gene. Genetic HU is rare but often overlooked entity. This study demonstrates, for the first time, the co-occurrence of renal glucosuria and familial renal HU, as well as the coexistence of xanthinuria type 2 and renal glucosuria in two patients. Rare FTL variants have not been previously reported in the literature as a possible cause of renal HU. Our study expands the spectrum of genetic findings of HU. Based on our findings, genetic studies have high diagnostic accuracy and should be used in patients with HU. Additionally, they may identify new candidate genes.

MicroRNAs (miRNAs) are small, non-coding RNAs that regulate the expression level of the target genes in the cell. Breast cancer is responsible for the majority of cancer-related deaths among women globally. It has been proven that deregulated miRNAs may play an essential role in the progression of breast cancer. It has been shown in many cancers, including breast cancer, that aberrant expression of miRNAs may be associated with drug resistance. This study investigated the effect of let-7b-5p, detected by bioinformatics methods, on Dox resistance through the Aurora Kinase B (AURKB) gene. In silico analysis using publicly available miRNA expression, GEO datasets revealed that let-7b-5p significantly downregulated in BC. Further in silico studies revealed that of the genes among the potential targets of let-7b-5p, AURKB was the most negatively correlated and may be closely associated with Dox resistance. Expression analysis via quantitative PCR confirmed that let-7b-5p was downregulated and AURKB was upregulated in breast cancer tissue samples. Later, functional studies conducted with MCF-10A, MCF-7, and MDA-MB-231 cell lines demonstrated that let-7b-5p inhibits cancer cells through AURKB and sensitizes them to Dox resistance. In conclusion, it has been shown that the let-7b-5p/AURKB axis may be significant in breast cancer progression and the disruption in this axis may contribute to the trigger of Dox resistance.

Background and Objectives: Curcumin is a turmeric-derived polyphenol, and it has shown anticancer potential in various cancers, including breast cancer (BC). Nevertheless, the molecular mechanisms underlying its effects remain incompletely defined. Hsa_circ_0001946 (CDR1as) is a circular RNA (circRNA) that promotes tumor progression by competitively inhibiting microRNA-7-5p (miR-7-5p) in BC. This study investigated whether curcumin regulates the hsa_circ_0001946/miR-7-5p/target gene axis in BC progression. Materials and Methods: BC cell lines (MCF-7 and T47D) and a non-cancerous human mammary epithelial cell line (MCF-10A) were treated with curcumin or transfected with circ_0001946 siRNA or miR-7-5p mimic. Cell proliferation, migration, apoptosis, and protein expression were analyzed by CVDK-8 analysis, a wound healing assay, and flow cytometry, respectively. Also, protein expression levels were quantified via Western blotting. In vitro and in silico findings were further validated by analyzing tumor and adjacent normal tissues from 65 luminal BC patients. Results: Curcumin inhibited the proliferation and migration of MCF-7 and T47D cells in a dose-dependent manner. Knockdown of hsa_circ_0001946 or overexpression of miR-7-5p significantly suppressed proliferation and migration and enhanced apoptosis in BC cells compared to the negative controls. Curcumin treatment led to the knockdown of hsa_circ_0001946, the overexpression of miR-7-5p, and the downregulation of hsa_circ_0001946, CKS2, TOP2A, and PARP1, while it upregulating miR-7-5p. The Western blot confirmed reduced CKS2 protein levels after curcumin treatment. The expression of both hsa_circ_0001946 and CKS2 was significantly upregulated in tumor tissues compared to that of matched adjacent normal tissues, whereas that of miR-7-5p was markedly downregulated. Conclusions: This preliminary study shows that curcumin suppresses BC tumorigenesis by modulating the hsa_circ_0001946/miR-7-5p/target gene axis. While these findings suggest a novel regulatory pathway and potential therapeutic targets, further in vivo validation and clinical trials are required to determine the translational relevance of curcumin in BC therapy.

Objective: Acute myeloid leukemia (AML) demonstrates prognostic heterogeneity, and the underlying pathophysiology is still not fully elucidated. This study investigated how the tumor suppressor miR-145-5p contributes to AML heterogeneity and associated biological processes through its potential target oncogenes. Materials and Methods: HL-60 and NB4 AML cells were transfected with miR-145-5p mimics using LipofectamineTM 2000. Apoptosis was evaluated by caspase-3 activity in NB4 cells, while cell viability was determined using WST-8 assays in HL-60 and NB4 cells. Putative miR-145-5p targets were predicted using the miRTarBase (v9.0) and miRNET(v2.0) databases. Followingfurtherfiltration, PPI, KEGG, and GO analyses were performed using in silico approaches. Expression profiling of the identified genes was evaluated via quantitative real-time polymerase chain reaction (qRT-PCR), and the data were statistically compared. Results: After miR-145-5p transfection, cell survival markedly decreased in both cell lines, and caspase-3 levels markedly increased in NB4 cells (p<0.05). Among the putative genes CDK4, CDK6, KLF4, NRAS, IRS1, and eIF4E identified via in silico analysis, CDK4 and KLF4 showed significantly decreased expression in both cell lines (p<0.05 for both). Conclusion: The results demonstrate that miR-145-5p potentially regulates apoptosis and proliferation in AML via target genes, including CDK4 and KLF4. Further studies may reinforce the biomarker potential of miR-145-5p and facilitate the advancement of innovative diagnostic and therapeutic approaches for AML.

Objective: miR-638-5p is a crucial tumor suppressor miRNA in several cancer types including, Acute Myeloid Leukemia (AML). This study aimed to analyze the role of miR-638-5p and its potential target genes in HL-60 and NB4 acute promyelocytic leukemia cell lines using in vitro method. Method: After the miR-638-5p mimic transfection into AML cells, the effect on cell viability was examined by the WST-8 method, and the effect on apoptosis was measured via the Caspase-3 quantification method. In silico tools such as miRWalk, miRDB, and miRTarBase were used to select the possible target genes of miR-638-5p. The expression levels of selected genes were investigated by qRT-PCR. The overall survival (OS) rate of AML patients was explored via the BloodSpot database, the Enrichr tool was used for enrichment analysis, and correlation analysis was performed using the Correlation AnalyzeR tool. Results: Decreased proliferation and increased apoptosis were determined in miR-638-5p mimic transfected cells compared to the controls. MECP2, PIM1, MEF2C, PGK1, Aand SPAG1 genes were selected as the potential targets of miR-638-5p for in vitro study. PGK1 and PIM1 expression levels were significantly suppressed in cells transfected with the miR-638-5p mimic. The OS investigation revealed that overexpression of MECP2, MEF2C, and PGK1 does not affect the survival of AML patients; however, overexpression of SPAG1 and PIM1 has a detrimental effect on AML survival. Also, a positive correlation was detected between PIM1 and PGK1 genes via enrichment analysis. Conclusions: miR-638-5p may contribute to AML pathogenesis by targeting the PGK1 and PIM1 genes, and this situation may indicate its potential as a biomolecule for regulating cell proliferation in AML cells.

Focal Segmental Glomerulosclerosis (FSGS) is a clinicopathological illness characterized by podocyte damage, impairing glomerular filtration, and substantial proteinuria, which often results in end-stage renal disease (ESRD). Divided into primary, secondary, genetic, and idiopathic categories, its diverse origin highlights the intricacy of its diagnosis and treatment. The existing dependence on immunosuppressive medicines highlights their side effects and inconsistent efficacy, underscoring the pressing necessity for innovative, focused treatments. Recent advancements in genomics and molecular biology have shown the significant involvement of genetic alterations, especially in podocyte-associated proteins, in the pathogenesis of FSGS. Identifying possible novel biomarkers for diagnosing FSGS and monitoring disease activity has revitalized interest in this condition. Recent data underscores the significance of non-coding RNAs, including microRNAs (miRNAs), circular RNAs (circRNAs), and long non-coding RNAs (lncRNAs), in the modulation of gene expression and podocyte functionality. Please check and confirm that the authors and their respective affiliations have been correctly identified and amend if necessary. Particular dysregulated miRNAs and circRNAs have demonstrated potential as biomarkers for early diagnosis and disease monitoring. Furthermore, understanding lncRNA-mediated pathways provides novel therapeutic targets. This review consolidates current progress in elucidating the genetic and molecular processes of FSGS, emphasizing biomarker identification and treatment innovation.

Objectives Curcumin plays a leading role as an epigenetic regulator in cancer. miR-15a-5p is a crucial non-coding RNA for breast cancer (BRCA) and various cancers due to its tumor suppressor role. In our study, we aimed to examine the curcumin/miR-15a-5p/target gene interaction in BRCA cells.Methods The effects of curcumin and miR-15a-5p on cell viability in the MCF7 cells were examined using the WST8 technique. The cell migration was determined using scratch wound assay. miR-15a-5p level was detected in curcumin-treated cells and miR-15a-5p transfected cells compared to control groups by RT-qPCR. Overexpressed genes in BRCA were found by bioinformatics tools (GSE41970 and TCGA). miR15a-5p potential target genes in the miRNet tool were selected in overlapped genes between GSE41970 and TCGA. Survival analysis of the selected genes was examined using the GEPIA2 tool. Relative expression levels of four selected genes were examined via qPCR.Results Cell viability and scratch-wound closure rate were reduced in curcumin-treated and miR-15a-5p mimic transfected MCF7 cells. miR15a-5p overexpressed in curcumin-treated and miR-15a-5p transfected cells. Eighty-three dysregulated upregulated genes were detected (in GSE41970 and TCGA). Among the possible target genes of miR-15a-5p in the miRNet tool, 10 upregulated genes were detected overlapping with GSE41970 and TCGA. CCNE1 and CHEK1 genes were found to be important for survival in BRCA. CCNE1 and BMI1 were decreased in curcumin-treated and miR-15a-5p transfected cells.Conclusions Curcumin treatment increased miR-15a-5p and downregulated selected target genes. Curcumin/miR-15a-5p interaction may be a much stronger negative regulator of the CCNE1 and BMI1 genes in BRCA.

Multiple myeloma (MM) is a malignant disease that causes abnormal immunoglobulin synthesis by bone marrow plasma cells. The relationship between MM and the TP53 pathway has not been fully elucidated in the literature. Investigation of the effect of the expression of genes in the TP53 pathway on the molecular pathogenesis and prognosis of multiple myeloma disease. We assessed the expression of 18 genes in the TP53 pathway in 48 MM patients and 31 healthy subjects by RT2-profiler PCR array technique, and investigated their possible association with the presence of cytogenetic aberrations. Twelve of the 18 genes ( APAF1, ATM, BAX, CASP9, CDK4, CDKN1A, CDKN2A, E2F1, MCL1, MDM2, MDM4, PTEN) ) expression levels were found to be statistically up-regulated in MM patients compared to controls. The CDK4, CDKN1A and MCL1 genes were found to have remarkable diagnostic power distinguishing MM and healthy controls (AUC=0.89;AUC=0.86;AUC=0.77, respectively and p<0.001 for all three) via using Receiver operating characteristic (ROC) analysis. Overexpression of CDK4 and CDKN1A, , which are involved in the cell cycle, and MCL1, , which is an important gene in the anti-apoptotic process, were found to be excessively increased in MM patients compared to controls in terms of mRNA fold change. In addition, the high sensitivity of these genes found in the ROC analysis results suggests that they may be suggested as potential biomarkers for MM.

Background Breast cancer (BC) is the most common cancer in women worldwide. Curcumin is a polyphenolic turmeric-derived compound that has anti-proliferative and anti-tumor properties in different cancer types by acting on multiple molecules. Circular RNAs (circRNAs) are non-coding, single-stranded, covalently closed RNA molecules that act as regulators of the microRNA (miRNA) activity. Recent studies show that circRNAs are potential contributors to the onset and progression of many cancer types. CircRNA ciRS-7 acts as an oncogene and accelerates tumor progression by competitively suppressing miR-7-5p in BC. However, whether curcumin can regulate ciRS-7 to inhibit BC progression is still unclear. Method Breast Cancer cell lines (MCF-7 and T47D) and normal epithelial cell line (MCF-10A) were cultivated and treated with Curcumin (5μM, 10 μM, 20 μM) or DMSO as a control. Also, the cell lines were transfected with ciRS-7 siRNA, miR-7-5p mimic, and their non-targeted controls. The cell viability was detected by Cell Viability Detection Kit-8 (CVDK-8) using an ELISA plate reader. To detect cell migration, scratch assay was performed after 24hs of transfection and a phase contrast microscope was used to acquire images. The effect on apoptosis was detected by Annexin V-FITC Apoptosis Detection Kit using flow cytometry. Potential target genes of miR-7-5p were identified by searching for the overlapping genes in miRNet and miRTarBase v8 with the overexpressed genes in BC patient tissue samples in the TCGA database. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of ciRS-7, miR-7-5p, and the selected genes. qRT-PCR experiments were performed in duplicate and the 2−ΔΔCt method was used for relative quantitation analysis. Results Curcumin treatment, depending on the increased doses, decreased cell proliferation through inducing apoptosis in MCF-7 and T47D cancer cells. Curcumin's anti-proliferative effect was shown to be quite restricted on normal MCF-10A cells, as compared to MCF-7 and T47D cancer cells. In MCF-7 and T47D cells treated with curcumin, cell migration was dramatically inhibited. Curcumin has very little influence on the migration of MCF-10A cells. In both cancer cell group that was transfected with ciRS-7 siRNA and miR-7-5p mimic, it was observed that apoptosis was increased, proliferation was suppressed, and migration was decreased compared to its control groups. Moreover, the expression of ciRS-7 was found to be significantly decreased in the curcumin-treated group, while miR-7-5p was shown to be significantly higher. CKS2 gene one of the possible target genes of miR-7-5p that was identified by using in silico approaches, was also downregulated in curcumin-treated BC cell lines compared to its control group. ciRS-7 and CKS2 expression levels were found to be downregulated, whereas miR-7-5p expression level was found to be elevated in MCF-7 and T47D cells that were transfected with ciRS-7 siRNA. Additionally, CKS2 gene expression was found to be downregulated in miR-7-5p mimic transfected cells. Conclusion Curcumin is derived from the turmeric plant (Curcuma longa) and has been used since 3000 B.C. as a food additive. Nowadays, curcumin is one of the most essential anti-cancer substances that was examined. However, investigations on the influence of curcumin on the circRNA-miRNA-mRNA axis are scarce. Curcumin has been demonstrated to inhibit proliferation and induce apoptosis of BC cells via the ciRS-7/miR-7-5p/CKS2 axis in the present study. Keywords Breast Cancer, Curcumin, ciRS-7, miR-7-5p, CKS2 Citation Format: Asmaa Abuaisha, Murat Kaya, Ilknur Suer, Selman Emiroglu, Fahrunnisa Abanoz, Mustafa Tukenmez, Neslihan Cabıoğlu, Mahmut Muslumanoglu, Kivanc Cefle, Sukru Palanduz, Sukru Ozturk. Curcumin inhibits breast cancer cell proliferation by regulating ciRS-7/miR-7-5p/CKS2 axis [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-28-04.