The application of Multiple Instance Learning (MIL) for classifying Whole Slide Images (WSIs) has gained extensive use in recent years, primarily due to the high cost and time consumption associated with pixel-level annotation of WSIs, which is challenging to accomplish. The advancements in MIL for WSIs have predominantly concentrated on two fronts: the development of superior feature extractors (for instance, utilizing self-supervised learning for training feature extractors) and the formulation of enhanced instance aggregation strategies. Regrettably, the majority of the most advanced approaches have neglected phenotypic variances among instances when employing attention mechanisms. To capitalize on the disparities between instance tissues, we have introduced a phenotypic self-distillation approach to MIL. Our framework is composed of three components: i) a self-supervised feature extractor based on contrastive learning and a phenotype extractor pre-trained on the Kather100K dataset, which automatically provides 9-class tissue phenotype labels (e.g., tumor epithelium, stroma, lymphocytes) without requiring manual annotation, ii) the incorporation of a self-distillation loss between the features of instances and their phenotypes to augment the informational content of both perspectives, and iii) the aggregation of MIL instances for the final MSI prediction. The efficacy of this framework was evaluated on two datasets: the TCGA-CRC dataset was used for training and internal testing with a fixed 70%/30% split, while the Tangshan People's Hospital cohort served as an independent external validation set. On the TCGA-CRC dataset (n = 360; 65 MSI-H, 295 MSS), our model achieved an AUC of 0.8846 and an accuracy of 0.84, using a fixed 70%/30% train-test split. On the Tangshan People's Hospital dataset (n = 472; 56 MSI-H, 426 MSS), the model attained an AUC of 0.7258 and an accuracy of 0.70.

BACKGROUND:Nanog, a transcription factor, is involved in cancer initiation and progression. OBJECTIVES:To explore the potential regulatory mechanism of Nanog in gastric cancer. MATERIAL AND METHODS:Immunohistochemistry (IHC) was used to examine E-cadherin, N-cadherin, Nanog, and KDM5B in gastric cancer (Ca), metastatic lymph node cancer (L), and adjacent normal tissues (N). MTT, migration, and invasion assays were used to evaluate cell viability, migration, and invasion of gastric cancer cells SGC-7901 induced by transforming growth factor (TGF)-β1, or with Nanog overexpression/knockdown. Western blot was used to examine the relative protein expression of E-cadherin, N-cadherin, Nanog, and lysine-specific demethylase 5B (KDM5B) in cells. KDM5B was silenced by siRNA in cells, and E-cadherin, N-cadherin, and H3K4me3 were examined with western blot, while the interaction between Nanog and KDM5B was examined using a co-immunoprecipitation (Co-IP) assay. Furthermore, H3K4me3 protein expression was validated in N, Ca, and L tissues using IHC. RESULTS:E-cadherin was hypoexpressed, while N-cadherin, Nanog, and KDM5B were hyperexpressed in the Ca and L groups compared with the N group. TGF-β1 and Nanog enhanced cell viability, migration, and invasion by promoting epithelial-mesenchymal transition (EMT). KDM5B was identified to bind to Nanog and was positively regulated by Nanog, and functioned as a negative regulator of histone H3 Lys4 trimethylation (H3K4me3) in SGC-7901 cells. KDM5B knockdown inhibited cell invasion and EMT. The Nanog/KDM5B/H3K4me3 pathway contributes to the proliferation, migration, and invasion of SGC-7901 cells by inhibiting E-cadherin and enhancing N-cadherin. CONCLUSIONS:This study provides new insights into the regulatory role and mechanism of Nanog/KDM5B/H3K4me3 in gastric cancer, which might be potential biomarkers in the diagnosis and prognosis of gastric cancer.

M2 macrophages constitute a major stromal component of gastric cancer (GC) and actively contribute to tumor progression through angiogenesis, tissue remodeling, and epithelial–mesenchymal transition (EMT). Our previous study identified CD44 variant 9 (CD44v9) expression in cytokeratin-negative stromal compartments as a marker with diagnostic and prognostic significance in gastric adenocarcinoma (GA); however, its cellular localization within the tumor microenvironment and functional relevance remain unclear. In this study, public single-cell RNA sequencing datasets from the Human Protein Atlas were analyzed to characterize CD44 expression across gastric tissue cell populations. Multiplex immunofluorescence was performed to validate the spatial distribution of CD44v9-positive cells in clinical GA specimens. CD44v9 expression was predominantly enriched in macrophages and showed significant co-localization with stromal M1 macrophages (CD68+CD86+CD163−), M2 macrophages (CD68+CD86−CD163+), and regulatory T (Treg) cells (CD3+CD4+FOXP3+CD8−). Functionally, CD44v9 was overexpressed in THP-1-derived M2 macrophages through lentiviral transduction, and these modified cells retained their inherent M2 macrophage phenotype. CD44v9 overexpression significantly enhanced TNF-α secretion (P < 0.05), without affecting IL-6 production (P > 0.05). In a co-culture system, CD44v9 overexpression M2 macrophages markedly promoted GC cell proliferation, migration, and invasion. These pro-tumorigenic effects were accompanied by EMT induction, characterized by decreased E-cadherin and increased N-cadherin and Vimentin (all P < 0.05) expression, along with elevated MMP2 and MMP9 (P < 0.05) levels in AGS cells. Collectively, these findings demonstrate that CD44v9 expression in stromal M2 macrophages promotes a pro-tumorigenic phenotype, possibly through TNF-α-mediated tumor-stroma crosstalk, thereby promoting EMT and enhancing cancer cell aggressiveness. CD44v9 expressed by M2 macrophages may represent a promising biomarker and therapeutic target in GA development.

Gastric cancer (GC) is a highly lethal malignancy, seriously threatening people's physical health. Accurate screening of gastric cancer could improve the survival rate of patients. Therefore, exploring noninvasive and efficient cancer screening methods for gastric cancer is of great significance. In the past few years, exosomes have received much attention for their potential in disease diagnosis and treatment. Here, the aim of this study was to explore the detection of serum exosomes via surface-enhanced Raman spectroscopy (SERS) technique based on TiN-Ag@Ag sol composite substrate, and its potential application in gastric cancer diagnosis is evaluated. Exosomes were extracted from the serum of 31 GC patients and 31 healthy controls (HC) using an exosome kit. This study used various machine learning algorithms such as principal component analysis linear discriminant analysis (PCA-LDA), partial least squares discriminant analysis (PLS-DA), support vector machine (SVM), and k-nearest neighbor (KNN) algorithm to analyze SERS spectra, in order to distinguish between HC and GC. The results show that the k-nearest neighbor algorithm performs the best in HC and GC classification. These results indicate that the combination of SERS and machine learning methods provides a new technological approach for gastric cancer screening. This study offers a new proposal for the universal applicability of analysis and identification with SERS of serum exosomes samples in clinical diagnosis.

ETHNOPHARMACOLOGICAL RELEVANCE:As a compound of traditional Chinese medicine (TCM), Bie Jia Jian pill (BJJP) is extensively used to treat the clinical chronic liver disease. Nevertheless, the specific mechanism through which BJJP affects hepatic fibrosis (HF) remains unknown. AIM OF THE STUDY:To explore the role and potential mechanism of BJJP involved in treating HF. MATERIALS AND METHODS:HF model of Sprague-Dawley (SD) rats was induced by a bile duct ligation (BDL). The function of BJJP involved in the intestinal microbiota (IM) and its metabolites in BDL-induced HF rats were explored through the 16S rRNA sequencing and untargeted metabolomics technologies. Network pharmacology was used to forecast mechanism underlying BJJP's anti-HF effects, which were validated in BDL-induced rats and trimethylamine N-oxide (TMAO)-induced LX-2 and HSC-T6 cells. RESULTS:BJJP effectively ameliorated pathological liver damage, inflammation, and fibrosis of the BDL-induced HF rats. BJJP regulated IM diversity and composition and interfered with trimethylamine (TMA)-flavin monooxygenase 3 (FMO3)-TMAO process. In vitro, BJJP significantly inhibited the TMAO-induced activation of hepatic stellate cells (HSCs) (rat HSC cell line, HSC-T6; human HSC cell line, LX-2). Network pharmacology results demonstrated that PI3K/AKT signal pathway is crucially involved in BJJP treatment of HF. Further research revealed that BJJP inhibited the PI3K/AKT signal pathway in BDL-induced HF rats. Moreover, TMAO activated the PI3K/AKT pathway, whereas BJJP suppressed TMAO-induced activation. Subsequent intervention with 740Y-P (the PI3K agonist) successfully neutralized the repression effect on PI3K/AKT signal pathway by BJJP. CONCLUSION:These results clearly show that BJJP attenuates HF by regulating the IM, as well as inhibiting PI3K/AKT pathway mediated by TMAO.

Among gynecologic malignancies, ovarian cancer is the most lethal, primarily due to its insidious early symptoms, lack of effective screening methods, and high risk of recurrence. It poses substantial challenges to clinical diagnosis and treatment. In recent years, the clinical application of poly ADP-ribose polymerase inhibitors has promoted a comprehensive management model that integrates targeted therapy with conventional treatments. This review, aiming to provide new perspectives and approaches for future research, summarizes the high-risk factors and first-line treatment strategies for ovarian cancer. Further studies should focus on optimizing personalized treatment strategies and exploring novel targeted therapies to improve patient survival outcomes.

The interaction between intratumoral microbiome and the tumor microenvironment (TME) has furthered our understanding of tumor ecology. Yet, the implications of their interaction for lung cancer management remain unclear. In the current work, we collected host transcriptome samples and matched intratumoral microbiome samples, as well as detailed clinical metadata from The Cancer Genome Atlas (TCGA) of 478 patients with lung adenocarcinoma (LUAD). Utilizing the multiomics integration approach, we comprehensively investigated the crosstalk between the TME and intratumoral microbiome in patients with LUAD. First, we developed a prognostic model based on the TME signatures (TMEindex) that clearly distinguished clinical, survival, and response to immunotherapy of patients with LUAD. Additionally, we found profound differences in intratumoral microbiota signatures, including alpha- and beta-diversity, among patients with different survival risks based on the TME signatures. In depth, we detected that genera Luteibacter and Chryseobacterium were strongly negatively and positively associated with patients’ survival risk, respectively, suggesting their opposing roles in cancer progression. Moreover, we developed a model that fused intratumoral microbial abundance information with TME signatures, called intratumoral microbiome-modified TMEindex (IMTMEindex), leading in predicting patient overall survival at 1-, 3-, and 5-years. Future clinical profiling of the specific intratumoral microbes in the TME could improve prognosis, inform immunotherapy, and facilitate the development of novel therapeutics for LUAD.

Accurate identification of clonal relationships between cell populations is crucial for investigating cellular differentiation trajectories and gaining insights into the underlying mechanisms of cancer initiation and development. The Single Cell Lineage Atlas (ScLineageAtlas; https://www.scladb.geneis.org.cn) is a comprehensive single-cell genomics database that characterizes cellular clones across various cancer types. The database currently includes 24 processed single-cell RNA sequencing datasets spanning 13 different cancer types. ScLineageAtlas leverages advanced computational methods to identify cellular clones, providing researchers with a detailed understanding of clone relationships and evolutionary dynamics. Additionally, the database offers comprehensive metadata for each sample, enabling researchers to explore contextual information and sample characteristics. The spatial visualization of cell clones presented in the ScLineageAtlas provides a valuable tool for enhancing our understanding of the genetic heterogeneity within the tumour microenvironment. Through the analysis of biological differences between these diverse cell populations, researchers can explore key genes and signalling pathways associated with cancer initiation, development, and therapeutic efficacy. In summary, the ScLineageAtlas serves as a user-friendly platform for data operations on cellular clones, facilitating the understanding of tumour heterogeneity, differentiation trajectories, and evolution. It thus contributes significantly to cancer research and clinical practice.

Background Metastasis-associated in colon cancer-1 (MACC1) is a novel oncogene involved in the growth and metastasis of tumors, which is overexpression in various tumors. MACC1 could promote the growth, invasion, and metastasis of colorectal cancer (CRC) by activating the HGF/MET signaling pathway in vivo . It has been confirmed that MACC1 mainly promotes the Warburg effect in gastric cancer cells through the PI3 K/AKT signaling pathway. Objective Here, we mainly investigated the association between MACC1 and the glycolysis process in CRC cells. Methods The expression of MACC1 in CRC and its relationship with the patient's survival were analyzed by TCGA database. We used different concentrations of glucose medium to culture HT-29 and HCT-116 with or without siMACC1. Cell Counting Kit-8 assay was used to detect cell proliferation. The content of lactic acid and glucose in cells was examined by enzyme-linked immunosorbent assay, and extracellular acidification rate was also determined with Seahorse XFe96 Extracellular Flux Analyzer. Western blot was used to detect the protein expressions related to glycolysis. Immunofluorescence was conducted to observe the expression and distribution of GLUT4. Results In this study, we observed that MACC1 was highly expressed in CRC and negatively correlated with the survival of patients. The expression of glycolysis related enzymes was significantly increased under the stimulation of different concentrations of glucose in HT-29 and HCT-116 cells. However, MACC1 knockdown could significantly reduce high glucose-induced the expressions of glycolysis-related enzymes. Besides, MACC1 knockdown could decrease the content of glucose and lactate, and inhibit glycolytic function in HT-29 and HCT-116 cells. Moreover, the expression of GLUT4 was significantly decreased in the two cell lines treated by 4.5 g/L or 9.0 g/L glucose with MACC1 knockdown. Additionally, MACC1 knockdown could inhibit high glucose-induced membrane translocation of GLUT4. Conclusions MACC1 knockdown can attenuate glucose metabolism by inhibiting the membrane translocation of GLUT4 in CRC cells.

TiN–Ag@Ag composite substrates were prepared via ammonia reduction nitridation followed by electrochemical deposition. Fabricated TiN–Ag@Ag substrates were characterized using X-ray diffraction, X-ray photoelectron spectroscopy, scanning electron microscopy, transmission electron microscopy, and ultraviolet-visible spectrophotometry. The surface-enhanced Raman spectroscopy activity of these substrates was evaluated using ibuprofen as the probe molecule. The size of the Ag particles prepared via electrochemical deposition was approximately 1 μm, and Ag nanoparticles with an average particle size of 100 nm were uniformly distributed on the surface of TiN–Ag films. The Raman signal of ibuprofen was significantly enhanced, and the minimum detection concentration of ibuprofen was 10–5 M. The mechanism by which the TiN–Ag@Ag composite substrate enhanced the Raman signals was analyzed using ultraviolet photoelectron spectroscopy and density functional theory implemented in the Gaussian software. Overall, charge transfer and the local electromagnetic field effect enhanced the Raman signals of ibuprofen.

The specificity and sensitivity of the current diagnostic and prognostic biomarkers for gastric cancer (GC) are limited. The present study aimed to evaluate the diagnostic and prognostic significance of cluster-of-differentiation gene 44 variant isoform 9 (CD44v9) and T cell immunoglobulin and mucin domain-containing protein 3 (TIM3) expression levels alone or combined in the tumor tissues of patients with GC and reveal the roles of CD44v9 and TIM3 in the cytokeratin (CK)(+) and CK- regions. Multiplex immunofluorescence staining was performed for CD44v9, TIM3 and CK using a tissue microarray. The tissues were divided into three regions based on CK expression: Total, CK+, and CK- regions. The diagnostic and prognostic value was evaluated using receiver operating characteristic curves, Kaplan-Meier and Cox regression analyses. The results demonstrated that the density of cells expressing CD44v9, TIM3 and co-expressing CD44v9 and TIM3 (CD44v9/TIM3) in both the CK+ and CK- regions of tumor tissues was significantly higher than those in normal tissues (P<0.001). Moreover, the expression of CD44v9 in the CK- region was significantly positively correlated with age and tumor grade (P<0.05), and the expression of CD44v9/TIM3 in the CK- region of tumor tissues was significantly positively correlated with age, tumor grade and metastasis (P<0.05). Furthermore, the area under the curve for TIM3 expression in the CK+ region was 0.709, with a sensitivity of 45.83% and a specificity of 85.54% (P<0.001). High expression of CD44v9 in the CK- region was also significantly associated with poor survival and independently predicted a poor prognosis in patients with GC (hazard ratio, 2.387; 95% confidence interval, 1.384-4.118; P<0.01). In conclusion, dividing tissue regions based on CK expression is important for the diagnosis of GC. The expression of TIM3 in the CK+ region demonstrated diagnostic potential for GC, and high expression of CD44v9 in the CK- region was an independent prognostic risk factor for patients with GC.

Abstract Purpose This study explored the relationship between lung function and secondary thrombocytopenia by analyzing Mendelian randomization (MR). Methods As instrumental genetic variables, appropriate single nucleotide polymorphisms (SNPs) were extracted from genome-wide association study (GWAS) data on lung function. We choose secondary thrombocytopenia as the outcome. Inverse variance weighting, weighted median methods, and MR- Egger's method were used to explore the association between lung function and the risk of secondary thrombocytopenia. The analysis results were shown as odds ratio (OR) and 95% confidence interval to evaluate the causal association between lung function parameters and secondary thrombocytopenia. Results The inverse variance weighted (IVW) analysis showed that forced expiratory volume in the first second (FEV1) significantly led to the occurrence of secondary thrombocytopenia [OR: 0.2126; 95% confidence interval (CI): 0.0508–0.8886, P = 0.0339], reduced peak expiratory flow (PEF) increased the occurrence of secondary thrombocytopenia [OR: 0.1018; 95% CI: 0.0143–0.7234, P = 0.0224]. The decline of FEV1 and PEF contributed to the risk of secondary thrombocytopenia. Furthermore, FVC (Forced Vital Capacity), FEV1/FVC, and lung volume were not associated with secondary thrombocytopenia. Conclusion FEV1 and PEF could be important precipitating factors of secondary thrombocytopenia, which provided new ideas for treating and preventing secondary thrombocytopenia.

Exosomes belong to a subpopulation of EVs that carry different functional molecular cargoes, including proteins, nucleic acids, metabolites, and lipids. Notably, evidence has demonstrated that exosomes participate in bidirectional cell–cell communication and act as critical molecular vehicles in regulating numerous physiological and pathological processes. Since the specific contents within exosomes carry the information from their cells of origin, this property permits exosomes to act as valuable biomarkers. This chapter summarizes the potential use of exosome components in diagnosing, prognosis, or monitoring and treating multiple cancers and other non-neoplastic diseases. We also discuss the deficiency of basic applications, including the limitations of research methods and different research institutions and the differences generated by specimen sources. Thus, a better understanding of the problem of exosome detection may pave the way to promising exosome-based clinical applications.

以制备的MOF@TiN-Ag/银溶胶复合基底为表面增强拉曼光谱(SERS)活性基底,对茶碱进行SERS检测,探讨了基于该复合基底的表面增强拉曼技术在药物检测方面的应用.首先,利用电化学阳极氧化结合氨气还原氮化法制备了氮化钛纳米管阵列,随后通过电化学沉积法制备TiN-Ag复合基底,并在其表面原位生长金属有机框架(MOF)包覆层得到MOF@TiN-Ag复合基底,将茶碱与银溶胶混合后滴加在该复合基底上进行表面增强拉曼光谱检测.结果表明,MOF@TiN-Ag/银溶胶复合基底中存在面心立方晶型TiN、金属单质Ag和MOF钴基3种物相;扫描电镜结果显示,TiN纳米管排列整齐,Ag纳米结构呈树枝状均匀分散在其表面;作为隔绝层的MOF粒子形状规整,覆盖在TiN-Ag表面;银溶胶纳米粒子呈圆球状分布在MOF@TiN-Ag复合基底表面.由于银纳米粒子与TiN-Ag复合基底可发生协同增强作用,加之MOF的富集特性,使得该复合基底具有优异的SERS性能,其对茶碱溶液的SERS检出限为1×10-5 mol/L,检测性能良好.所制备的MOF@TiN-Ag/银溶胶复合基底拓宽了SERS在药物检测领域的应用.

Exosomes have been implicated in inflammation-related diseases, such as hepatic fibrosis (HF) and renal fibrosis, via transferring bioactive cargoes to recipient cells. This study aimed to investigate the possible effect of hepatic stellate cell (HSC)-derived exosomes on the initiation and development of HF by delivering microRNA (miR)-199a-5p. In HF rats with cholestasis induced by ligating the common bile duct, miR-199a-5p was upregulated while SIRT1 was downregulated in liver tissues from bile duct ligation (BDL) rats compared with that of sham rats. Furthermore, miR-199a-5p expression was upregulated, but the mRNA and protein expression levels of SIRT1 were downregulated in TGF-β1-activated LX-2. miR-199a-5p promoted the proliferation and further activation of LX-2 and enhanced the expression levels of the HF markers COL1A1 and α-SMA. Subsequently, the binding of miR-199a-5p to the 3'UTR of SIRT1 mRNA was predicted by bioinformatics websites and evidenced by fluorescent reporter assay. Knocking down SIRT1 enhanced the abilities of LX-2 cell proliferation, migration, and colony formation and increased the expression levels of the HF markers α-SMA and COL1A1. LX-2-derived exosomal miR-199a-5p transferred to LX-2 and THLE-2, inhibited the proliferation of THLE-2, and promoted the epithelial mesenchymal transition (EMT) and senescence of THLE-2. Furthermore, in vivo results suggested that miR-199a-5p overexpression aggravated HF in BDL rats; increased miR-199a-5p, α-SMA, and COL1A1 expression levels; and significantly upregulated the serum ALT, AST, TBA, and TBIL levels. However, reverse results were obtained with inhibited miR-199a-5p expression. In conclusion, HSC-derived exosomal miR-199a-5p may promote HF by accelerating HSC activation and hepatocyte EMT by targeting SIRT1, suggesting that miR-199a-5p and SIRT1 may serve as potential therapeutic targets for HF.

Objective:To investigate the effects and mechanisms of zinc finger E-box binding homeobox transcription factor-2 ( ZEB2) on the proliferation, colony formation, migration, and invasion abilities and the epithelial-mesenchymal transition (EMT) of PANC-1 cells, a human pancreatic cancer cell line.Methods:Data on the expression of ZEB2 in pancreatic cancer tissues and paracancerous tissues from The Cancer Genome Atlas (TCGA) database were analyzed. PANC-1 pancreatic cancer cells were divided into si-NC group, si- ZEB2 group, pcDNA3.1 group, and pcDNA3.1- ZEB2 group. qRT-PCR and Western blot were conducted to confirm the effectiveness of ZEB2 knockdown or overexpression. CCK-8, colony formation, wound healing, and Transwell assays were conducted to examine the effects of ZEB2 on the proliferation, colony formation, migration, and invasion of PANC-1 cells. qRT-PCR and immunofluorescence assays were performed to examine the expression of E-cadherin and vimentin, the EMT markers, in the cells. Prediction of proteins interacting with ZEB2 was made through the STRING database.Results:TCGA database analysis showed that the expression level of ZEB2 in pancreatic cancer tissues was significantly higher than that in adjacent tissues ( P<0.05). Compared with those of cells in the control group, the proliferation, colony formation, migration, and invasion of cells in the si- ZEB2 group were decreased ( P<0.05). Compared with those of cells in the pcDNA3.1 group, the proliferation, colony formation, migration and invasion of cells in the pcDNA3.1- ZEB2 group were increased (all P<0.05). According to the results of qRT-PCR and immunofluorescence assays, compared with those of the si-NC group, the expression of E-cadherin mRNA, an epithelial marker, in the si- ZEB2 group increased, while the expression of vimentin mRNA, an mesenchymal marker, and the protein decreased. Compared with those of the pcDNA3.1 group, the expression of E-cadherin mRNA in the PANC-1 cells of the pcDNA3.1- ZEB2 group decreased, while the expression of vimentin mRNA and the protein increased (all P<0.05). Analysis with the STRING database predicted that 10 proteins had close interaction with ZEB2.Conclusion:Overexpression of ZEB2 promotes the migration, invasion, and the EMT process of PANC-1 pancreatic cancer cells.

严重急性呼吸综合症冠状病毒2(seveve acute respiratiory syndrome coronavivus 2,SARS-CoV-2)具有极高传染性,对人类生命安全造成极大威胁.SARS-CoV-2的早期和快速诊断在抗击疫情方面发挥着重要作用,需要一种快速、灵敏且廉价的检测系统.表面增强拉曼光谱(surface-enhanced raman scattering,SERS)作为一种前沿分析技术具备指纹识别能力,可提供具有高特异性、灵敏度和多重检测的定量结果,在病毒的即时诊断方面具有广阔前景,目前已被应用于多种甲型流感病毒亚型的检测.侧流免疫分析(lateral flow immunoassay,LFIA)等床旁(point of care,POC)设备提供了快速、高效、可靠的平台.然而,LFIA在SARS-CoV-2检测的定量和灵敏分析方面存在局限性.为了解决这些问题,本文综述了 SERS技术与LFIA结合,用于SARS-CoV-2的定量分析现状及未来挑战.

本研究旨在揭示miR-192-5p对胰腺癌(pancreatic cancer,PC)细胞增殖、迁移和侵袭的影响,为胰腺癌的临床诊断和治疗提供实验依据.采用实时荧光定量PCR(real time quantitative PCR,RT-qPCR)检测miR-192-5p在人正常胰腺上皮细胞hTERT-HPNE和胰腺癌细胞系AsPC-1中的表达水平;利用Cell Counting Kit-8(CCK-8)、集落形成、划痕以及Transwell实验检测miR-192-5p对AsPC-1细胞增殖、克隆形成、迁移和侵袭能力的影响;生物信息学方法预测miR-192-5p的候选靶基因,并通过双萤光素酶报告实验对miR-192-5p的靶基因进行验证;RT-qPCR 检测 miR-192-5p 对 E 盒锌指结合同源框 2(Zinc finger E-box binding homeobox 2,ZEB2)mRNA表达的影响.结果显示,miR-192-5p在胰腺癌细胞系AsPC-1中的表达水平明显低于正常胰腺上皮细胞hTERT-HPNE中的;胰腺癌细胞系AsPC-1中转染miR-192-5p mimics后其miR-192-5p的表达水平显著升高,并能抑制ZEB2 mRNA的表达,抑制胰腺癌细胞系AsPC-1的增殖、集落形成、迁移和侵袭;胰腺癌细胞系AsPC-1中转染miR-192-5p inhibitor后其miR-192-5p的表达显著降低,促进ZEB2 mRNA的表达,促进胰腺癌细胞系AsPC-1的增殖、集落形成、迁移和侵袭;双萤光素酶报告实验表明ZEB2是miR-192-5p的功能靶基因.本研究结果提示miR-192-5p可能通过靶向ZEB2而抑制胰腺癌细胞系AsPC-1的增殖、迁移和侵袭.