Background Tumor metastasis is the primary cause of poor survival in gastric cancer (GC) patients. PVT1 and EIF4A1 have been shown to synergistically promote GC metastasis and invasion. However, the underlying mechanism remains unclear. Methods The binding site between PVT1 and EIF4A1 was identified by RNA-binding protein immunoprecipitation (RIP). 4D-DIA proteomics combined with mass spectrometry was used to identify downstream proteins regulated by PVT1 and EIF4A1. From the differentially expressed proteins, Stanniocalcin-1 ( STC1) expression was validated in GC tissues, cell lines, and animal metastatic tumor samples. Following STC1 knockdown, cell proliferation and migration were assessed using MTT, colony formation, wound healing, and Transwell assays. Epithelial-mesenchymal transition (EMT) and Notch1 signaling were also evaluated. Results PVT1 bound to EIF4A1 through its N-terminal domain (NTD). Proteomic analysis revealed that PVT1 and EIF4A1 jointly upregulate STC1 expression. Immunohistochemistry (IHC) on a tissue microarray containing 183 GC samples, 128 adjacent normal tissues, and animal metastatic lesions demonstrated that STC1 was significantly upregulated in GC and correlated with metastasis, histological type, and tumor stage. Inhibition of EIF4A1 reduced STC1 expression, suggesting that STC1 is a key downstream target of the PVT1/EIF4A1 axis. Knockdown of STC1 attenuated GC cell migration and proliferation, and suppressed both EMT and the Notch1 signaling pathway. Conclusion STC1 acts as a critical downstream effector of PVT1 and EIF4A1, promoting GC cell metastasis and proliferation, thereby driving malignant progression. This mechanism involves the regulation of the Notch1 signaling pathway and the EMT process.

OBJECTIVE:This systematic review aims to evaluate the extensive application of platelet-rich plasma (PRP) as a novel autologous cell therapy across various clinical disciplines in recent years. METHOD:A comprehensive search was conducted across PubMed, Scopus, Embase, and the Cochrane Library from 2009 to 2024, resulting in the identification of 153 research articles. The inclusion criteria were focused on studies that reported on the molecular mechanisms of action and clinical efficacy. The data were synthesized through qualitative narrative analysis and thematic classification. RESULT:Key active components found in PRP, including platelet-derived growth factor, transforming growth factor-β, and vascular endothelial growth factor, exhibit regulatory effects on fibroblasts, endothelial cells, and immune cells. These components facilitate cell proliferation and migration via signaling pathways such as PI3 K/Akt and MAPK/ERK. Notably, differences exist between the use of PRP alone versus its application in combination therapies for conditions such as bone and ligament repair, female infertility, alopecia, facial rejuvenation, diabetic foot ulcers, and fistulas. Factors such as PRP storage methods, white blood cell concentration levels, platelet concentration variations, and whether it is activated ex vivo can significantly influence therapeutic outcomes. CONCLUSION:PRP represents a highly promising autologous treatment modality. Standardizing protocols for PRP preparation alongside disease-specific treatment strategies will enhance precision in clinical applications.

We investigated the correlation between bromodomain-containing protein 4 (BRD4) and HPV16 viral load in cervical squamous cell carcinoma and non-cancerous tissues, as well as the effects of BRD4 degradation regent MZ1 on the viral load, proliferation and migration of cervical cancer cells. Real-time fluorescence quantitative PCR showed that the viral load of cervical cancer specimens was significantly higher than that of non-cancer specimens. Immunohistochemical assay showed that BRD4 expression was elevated in cervical cancer specimens (P < 0.001) and in specimens with high viral load (P < 0.0001). Treatment of cervical cancer cells of SiHa, HeLa and CaSki with BRD4 degradation regent MZ1 significantly reduced viral load and inhibited cell proliferation and migration. Nude mouse xenograft tumor confirmed that the tumor volume, and tumor weight of the MZ1-treated mice were significantly lower than those of the control group. Expression of BRD4 and cell proliferation molecule Ki67 in the tumor sections of MZ1-treated mice was significantly decreased, while apoptosis molecule cleaved caspase-3 expression was significantly increased. Moreover, the viral load in the MZ1-treated group was significantly lower than that in the control group. These findings suggest that BRD4 has a potential role in HPV16 viral replication, and MZ1 has a favorable effect in inhibiting viral replication, increasing apoptosis and suppressing the proliferation and migration of cervical cancer cells.

The infection by Kaposi's sarcoma-associated herpesvirus (KSHV) is one of the most common causes of death in AIDS patients. Our studies have found that KSHV can infect SH-SY5Y cells (named SK-RG) in vivo and mTOR was up-regulated, which results in remarkable enhancement of cell proliferation, migration. But the regulatory role of mTOR in KSHV infected neurons has not yet been fully elucidated. Here, we find that miR-769-3p is decreased in SK-RG cells, which can exert anti-KSHV effect through negatively regulating the expression of mTOR. The knockdown of mTOR or overexpress of miR-769-3p decreased the proliferation, migration ability and cell cycle related protein of SK-RG cells, and the expression of KSHV related genes. In contrast, activating mTOR function by 3BDO treatment weakened the cellular behaviors of miR-769-3p overexpressing cells. Meanwhile, overexpressed miR-769-3p and rapamycin showed a shared inhibition trend in the effects on cell proliferation and motility. Our data indicated that miR-769-3p can inhibit cell proliferation and migration by down regulating mTOR in KSHV infected SH-SY5Y cells, and can be a candidate molecule for anti-KSHV therapy.

Background:: This study aimed to investigate the influence of Notch1 on c-Fos and the effect of c-Fos on the proliferation of Kaposi's sarcoma-associated herpesvirus (KSHV)-infected neuronal cells. Methods:: Real-time PCR and western blotting were used to determine c-Fos expression levels in KSHV-infected (SK-RG) and uninfected SH-SY5Y cells. C-Fos levels were measured again in SK-RG cells with or without Notch1 knockdown. Next, we measured c-Fos and p-c-Fos concentrations after treatment with the Notch1 γ-secretase inhibitor LY-411575 and the Notch1 activator Jagged-1. MTT and Ki-67 staining were used to evaluate the proliferation ability of cells after c-Fos levels downregulation. CyclinD1, CDK6, and CDK4 expression levels and cell cycle were analyzed by western blotting and flow cytometry, respectively. After the c-Fos intervention, the KSHV copy number and gene expression of RTA, LANA and K8.1 were analyzed by real-time TaqMan PCR. Results:: C-Fos was up-regulated in KSHV-infected SK-RG cells. However, the siRNA-mediated knockdown of Notch1 resulted in a significant decrease in the levels of c-Fos and p-c-Fos (P <0.01, P <0.001). Additionally, a decrease in Cyclin D1, CDK6, and CDK4 was also detected. The Notch1 inhibitor LY-411575 showed the potential to down-regulate the levels of c-Fos and p-c-Fos, which was consistent with Notch1 knockdown group (P <0.01), whereas the expression and phosphorylation of c-Fos were remarkably up-regulated by treatment of Notch1 activator Jagged-1 (P <0.05). In addition, our data obtained by MTT and Ki-67 staining revealed that the c-Fos down-regulation led to a significant reduction in cell viability and proliferation of the SK-RG cells (P <0.001). Moreover, FACS analysis showed that the cell cycle was arrested in the G0/G1 stage, and the expressions of Cyclin D1, CDK6, and CDK4 were down-regulated in the c-Fos-knockdown SK-RG cells (P <0.05). Reduction in total KSHV copy number and expressions of viral genes (RTA, LANA and K8.1) were also detected in c-Fos down-regulated SK-RG cells (P <0.05). Conclusion:: Our findings strongly indicate that c-Fos plays a crucial role in the promotion of cell proliferation through Notch1 signaling in KSHV-infected cells. Furthermore, our results suggest that the inhibition of expression of key viral pathogenic proteins is likely involved in this process.

Objective:To explore and analyze the expression of eukaryotic translation elongation factor 1 alpha 2 (eEF1A2) gene in cervical cancer tissues, its relationship with patient survival, gene mutations, and changes in copy number in cervical cancer and chronic cervicitis tissues. Methods:The expression of the eEF1A2 gene in cervical cancer and its relationship with patient survival were analyzed using gene expression profile interactive analysis. Changes in eEF1A2 expression in cervical cancer tissues were analyzed using cBioPortal, a portal for cancer genomics analysis. The eEF1A2 copy number in cervical cancer tissues and chronic cervicitis tissues was determined by real-time fluorescence quantitative polymerase chain reaction. The relationship between the expression of eEF1A2 protein and the clinical stage, pathological grade, and patient survival of cervical cancer was analyzed by the database: The Human Protein Atlas, an integrated repository portal for tumor-immune system interactions. Results:Gene expression profile interactive analysis database analysis showed no significant differences in the expression of eEF1A2 between cervical cancer and normal cervical tissues (P > .05). The eEF1A2 gene expression level was not correlated with the survival of cervical cancer patients (P > .05). Analysis of the cBioPortal database showed that 18 of 297 cervical cancer patients had eEF1A2 gene changes, including missense mutation, splice mutation, amplification, and messenger RNA increase. There was no significant difference in eEF1A2 gene copy number between cervical cancer and chronic cervicitis (P > .05). The Human Protein Atlas and an integrated repository portal for tumor-immune system interactions database analysis of immunohistochemical data showed that eEF1A2 protein expression was no significant difference in clinical stage, pathological grade and patient survival of cervical cancer (P > .05). Conclusion:The eEF1A2 gene was mutated in cervical cancer tissues. The eEF1A2 gene copy number was not associated with changes in the expression of the eEF1A2 gene in cervical cancer tissues.

目的 探讨表皮生长因子样重复序列和盘状蛋白I样结构域3(EDIL3基因)在宫颈癌中表达及其对宫颈癌细胞SiHa、HeLa迁移的作用.方法 运用GEPIA、UALCAN、OncoLnc数据库分析EDIL3基因mRNA在宫颈癌组织中的表达水平、基因启动子甲基化状态和预后的意义;采用免疫组织化学染色、Western blot、qRT-PCR分别检测宫颈癌组织和细胞中EDIL3蛋白及EDIL3基因mRNA表达水平;构建EDIL3-pcDNA3.1过表达质粒,转染分为实验组(转染过表达EDIL3-pcDNA3.1质粒)和对照组NC(转染空质粒),transwell实验检测宫颈癌细胞SiHa、HeLa的迁移.结果 生物信息学数据库分析显示EDIL3基因mRNA在宫颈癌组织中低表达并且存在启动子高甲基化(P＜0.05),在宫颈癌不同分期中EDIL3基因的表达无差异(P＞0.05),EDIL3高表达与宫颈癌患者不良预后相关(P＜0.05);免疫组织化学染色、Western blot、qRT-PCR与数据库分析结果一致,在宫颈癌组织和细胞中EDIL3基因均低表达(P＜0.05),且其表达水平与HPV感染无明显相关性(P＞0.05);Transwell实验显示过表达EDIL3后增强宫颈癌细胞SiHa、HeLa的迁移能力(P＜0.05).结论 EDIL3基因在宫颈癌组织和细胞中低表达并不受HPV感染的影响,而EDIL3高表达影响患者预后且促进宫颈癌细胞SiHa、HeLa的迁移能力.

BACKGROUND:Cervical cancer is a common gynecological malignancy. Gene microarray found that TCP11 gene was highly expressed in cervical cancer. However, the effect of TCP11 gene on the proliferation, apoptosis and migration of cervical cancer cells and its underlying molecular mechanisms are unclear.METHODS:GEPIA database, tissue microarray, western blot and qRT-PCR were used to analyze the expression of TCP11 gene in cervical cancer tissues and cells and its relationship with patients' survival rate. The cell cycle and apoptosis were detected by flow cytometry, and the expressions of cell cycle and apoptosis related molecules and EMT-related molecules were detected by Western blot and qRT-PCR.RESULTS:The results showed that TCP11 gene was highly expressed in cervical cancer tissues and cells compared with normal cervical tissues and cells, and its expression was positively correlated with patients' survival rate. The results of proliferation and migration assays showed that TCP11 overexpression inhibited the proliferation and migration of HeLa and SiHa cells. The results showed that TCP11 overexpression blocked the cell cycle of HeLa and SiHa cells, decreased the expression of CDK1 and Cyclin B1, and increased the apoptosis and the expression of caspase-3, cleaved-caspase-3 and cleaved-PARP. TCP11 overexpression increased the protein and mRNA expression of EMT-related molecules ZO-1 and E-cadherin. Conversely, TCP11 knockdown promoted the proliferation of HeLa and SiHa cells and the migration of HeLa cells.CONCLUSIONS:TCP11 overexpression significantly inhibited the occurrence and development of cervical cancer cells, it may be a potentially beneficial biomarker for cervical cancer.

目的 探讨微小RNA-769-3p(miR-769-3p)对卡波氏肉瘤相关疱疹病毒(KSHV)感染的神经细胞SH-SY5 Y(SK-RG)增殖及迁移方面的作用.方法 将SK-RG细胞分为NC mimics转染组(转染NC mimics)、miRNA-769-3p mimics转染组(转染miRNA-769-3p mimics)、miRNA-6815-5p mimics转染组(转染miRNA-6815-5p mimics)、miRNA-6874-3p mimics 转染组(转染miRNA-6874-3p mimics)和miRNA-7974 mimics转染组(转染miRNA-7974 mimics).用实时荧光定量逆转录聚合酶链反应和蛋白质印迹法检测哺乳动物西罗莫司靶蛋白(mTOR)mRNA和蛋白的表达水平,用双荧光素酶报告基因实验检测miR-769-3p对mTOR的靶向调控,用噻唑蓝法和平板克隆实验检测细胞的增殖情况,用划痕实验和 Transwell 实验检测细胞的迁移情况.结果 SK-RG细胞中NC mimics转染组、miRNA-769-3p mimics转染组的mTOR蛋白相对表达水平分别为 1.00±0.06 和 0.60±0.04,细胞第 5 天的增殖率为4.15±0.14和3.36±0.17,克隆形成数分别为(82.67±3.51)和(53.00±3.61)个,相对迁移率分别为 1.00±0.01 和0.66±0.01,迁移细胞数分别为(174.30±5.86)和(131.00±5.29)个,miRNA-769-3p mimics转染组的上述指标与 NC mimics转染组比较,差异均有统计学意义(均P＜0.05).结论 miR-769-3p可能通过靶向抑制 mTOR 的表达而降低 SK-RG 细胞的增殖和迁移能力,提示miR-769-3p可能作为治疗KSHV感染的小分子核酸药物.

The disease caused by Kaposi's sarcoma-associated herpesvirus (KSHV) is one of the major causes of death of individuals with acquired immunodeficiency syndrome (AIDS). Development of anti-KSHV drugs is thus crucial. In this study, we investigated the effect of parthenolide (PTL) on the proliferation and NF-κB signaling pathway of KSHV-infected cells. iSLK.219 and KSHV-infected SH-SY5Y cells (SK-RG) were treated with PTL, TaqMan real-time quantitative PCR was used to determine the number of copies of the KSHV genome, and mRNA and protein expression of KSHV genes were analyzed by real-time PCR and immunocytochemistry. A cell viability test was used to measure cell proliferation, and flow cytometry was used to examine the effect of the drug on the cell cycle. Cyclin D1, CDK6, CDK4, and NF-κB-related proteins, including IKKβ, P-p65, and P-IKB-α, were detected by Western blot. The results showed that PTL altered the morphology of the cells, reduced the KSHV copy number, and suppressed the production of ORF50, K8.1, and v-GPCR mRNA and the LANA, ORF50, and K8.1 proteins. It blocked the G1 phase in iSLK.219 cells and decreased the levels of cyclin D1, CDK6, and CDK4 as well as the levels of NF-κB signaling proteins, including IKKβ, P-p65, and P-IKB-α. Together, these results suggest that PTL is a candidate drug that can decrease KSHV pathogenicity by suppressing cell proliferation and inhibiting the NF-κB signaling pathway in KSHV-infected cells.

Background Cervical cancer is currently estimated to be the fourth most common cancer among women worldwide and the leading cause of cancer-related deaths in some of the world’s poorest countries. C/EBPβ has tumor suppressor effects because it is necessary for oncogene-induced senescence. However, C/EBPβ also has an oncogenic role. The specific role of C/EBPβ in cervical cancer as a tumor suppressor or oncoprotein is unclear. Objective To explore the role of the C/EBPβ protein in cervical tumorigenesis and progression. Methods Quantitative RT-PCR was used to analyze C/EBPβ (15 cervical cancer tissue samples and 15 corresponding normal cervical tissue samples) , miR-661 , and MTA1 mRNA expression in clinical samples (10 cervical cancer tissue samples and 10 corresponding normal cervical tissue samples). Immunohistochemistry was used to analyze C/EBPβ (381 clinical samples), Ki67 (80 clinical samples) and PCNA ( 60 clinical samples) protein expression. MALDI-TOF MassARRAY was used to analyze C/EBPβ gene methylation (13 cervical cancer tissues and 13 corresponding normal cervical tissues). Cell proliferation was analyzed by CCK-8 in cervical cancer cell lines. Western blotting and immunohistochemistry were performed to detect C/EBPβ protein expression levels, and mRNA expression was analyzed by quantitative RT-PCR analysis. Flow cytometry was performed to measure cell cycle distribution and cell apoptosis. Colony formation, Transwell, cell invasion, and wound healing assays were performed to detect cell migration and invasion. Results C/EBPβ protein expression was significantly reduced in cervical cancer tissues compared with cervicitis tissues ( P < 0.01). Ki67 protein and PCNA protein expression levels were significantly higher in cervical cancer tissues compared with cervicitis tissues. The rate of C/EBPβ gene promoter methylation of CpG12, 13, 14 and CpG19 in cervical cancer tissues was significantly increased compared with normal cervical tissue ( P < 0.05). In addition, C/EBPβ was overexpressed in cervical cancer cells and this overexpression inhibited cell proliferation, migration, invasion, arrested cells in S phase, and promoted apoptosis. Conclusions We have demonstrated that C/EBPβ decreased in cervical cancer tissues and overexpression of the C/EBPβ gene in cervical cancer cells could inhibit proliferation, invasion and migration.

目的 探讨TCP11基因在宫颈癌组织和细胞中的表达水平以及与HPV感染、患者临床特征和预后之间的关系.方法 运用UALCAN数据库及GEPIA数据库分析TCP11基因在宫颈癌中的表达及其与宫颈癌患者年龄和肿瘤分级的关系;运用The Human Protein Atlas(HPA)数据库、UALCAN数据库及Gene Expression Profiling Interactive Analysis(GEPIA)数据库分析TCP11基因的表达与宫颈癌患者生存率的关系;运用UALCAN数据库分析在宫颈癌中TCP11基因启动子甲基化状态.运用组织芯片分析TCP11基因在正常宫颈及宫颈癌组织中表达水平.利用第二代杂交捕获技术(HC-2)检测所收集样本HPV感染情况.体外培养HaCaT细胞(一种永生化的上皮细胞)及宫颈癌SiHa、HeLa、C33A细胞,利用Western blot、qRT-PCR分别检测宫颈癌细胞中TCP11蛋白和mRNA表达水平.结果 运用HPA、UALCAN、GEPIA数据库分析发现TCP11基因在宫颈癌组织中高表达,且与宫颈癌患者生存率显著相关(P<0.05);TCP11基因表达与宫颈癌患者的年龄及肿瘤分级无显著相关性(P>0.05).组织芯片结果统计发现TCP11基因在宫颈癌组织中高表达(P<0.05).与HaCaT细胞相比,TCP11蛋白和mRNA在宫颈癌SiHa、HeLa、C33A细胞系中均显著升高(P<0.05).HC-2结果显示TCP11基因的表达可能不受HPV感染的影响(P>0.05).结论 TCP11基因在宫颈癌组织及细胞中高表达,其表达水平与患者预后相关且可能不受HPV感染的影响.

"蛋白质的结构与功能"是生物化学与分子生物学课程中第一部分内容,也是重要章节,在医学生的培养体系中具有重要意义.本文以教育部颁发的课程思政文件作为指导方针,并以"蛋白质的结构与功能"这一章节为例,围绕着政治认同、家国情怀、文化素养、宪法法治意识、道德修养五个方面重点优化该章节的思政内容.培养学生正确的世界观、人生观与价值观,促进思政教育与教学有机融合,为生物化学与分子生物学课程思政建设提供实践参考.

Background We aimed to investigate the effects of miR-34a-5p on c-fos regulation mediating the malignant behaviors of SH-SY5Y cells infected with Kaposi’s sarcoma-associated herpesvirus (KSHV). Methods The KSHV-infected (SK-RG) and uninfected SH-SY5Y parent cells were compared for differentially expressed miRNAs using transcriptome sequencing. Then miR-34a-5p was upregulated in SK-RG cells by the miRNA mimics transfection. Cell proliferation ability was determined by MTT and plate clone assays. The cell cycle was assessed by flow cytometry analysis, and CDK4, CDK6, cyclin D1 levels were determined by Western blot analysis. The migration behavior was detected by wound healing and transwell assays. The protein levels of MMP2 and MMP9 were measured by Western blot analysis. The regulation of c-fos by miR-34a-5p was detected by the dual-luciferase reporter gene assay. Rescue assays were carried out by upregulating c-fos in miR-34a-5p-overexpressing SK-RG cells. KSHV DNA copy numbers and relative virus gene expressions were detected. Xenograft tumor experiments and immunohistochemistry assays were further used to detect the effects of miR-34a-5p. Results miR-34a-5p was lower in SK-RG cells. Restoration of miR-34a-5p decreased cell proliferation and migration, leading to a G1 cell cycle arrest and down-regulation of CDK4/6, cyclin D1, MMP2, MMP9. KSHV copy number and expression of virus gene including latency-associated nuclear antigen (LANA), replication and transcription activator (RTA), open reading frame (K8.1), and KSHV G protein-coupled receptor (v-GPCR) were also reduced. Furthermore, c-fos is the target of miR-34a-5p, while enhanced c-fos weakened cellular behaviors of miR-34a-5p-overexpressing cells. Xenograft experiments and immunohistochemistry assays showed that miR-34a-5p inhibited tumor growth and virus gene expression. Conclusion Upregulated miR-34a-5p in KSHV-infected SH-SY5Y cells suppressed cell proliferation and migration through down-regulating c-fos. miR-34a-5p was a candidate molecular drug for KSHV-infected neuronal cells.

目的 研究干扰mTOR对人类疱疹病毒8(HHV-8)感染的神经细胞SH-SY5Y的增殖和迁移能力的影响.方法 通过Western blot实验和qRT-PCR法验证HHV-8感染的SH-SY5Y神经细胞的mTOR表达水平.采用Lipofectamine 2000分别将si-mTOR与si-NC转染进KSHV感染的SH-SY5Y细胞中,采用Western blot和qRT-PCR技术检测转染效率;qRT-PCR检测HHV-8病毒相关因子LANA、RTA、K8.1的mRNA表达水平;采用CCK-8法和划痕实验检测细胞增殖和迁移能力;利用PI染色结合流式细胞术检测细胞周期;采用Western blot检测周期相关蛋白cyclin D1和p27的表达.结果 Western blot和qRT-PCR结果显示HHV-8感染的SH-SY5Y细胞中mTOR的表达水平高于SH-SY5Y细胞(P＜0.05).在干扰mTOR的HHV-8感染的SH-SY5Y细胞中,HHV-8病毒相关因子LANA、RTA、K8.1的表达水平均降低(P＜0.05).CCK-8和划痕实验结果表明干扰mTOR组细胞增殖、迁移能力低于对照组(P＜0.05).流式细胞术结果提示干扰mTOR组的细胞周期阻滞在G0/G1期(P＜0.05).Western blot结果显示cyclin D1表达水平降低,p27表达水平升高(P＜0.05).结论 干扰mTOR能够降低HHV-8感染的SH-SY5Y细胞的增殖和迁移能力,并将其细胞周期阻滞在G0/G1期,针对mTOR的靶向调控在治疗HHV-8感染引起的神经系统疾病有一定的作用.

目的 观察真核翻译起始因子4A1(eukaryotic translation initiation factor 4A1,EIF4A1)在胃癌组织中的表达以及沉默EIF4A1对人胃癌细胞株MGC-803增殖、迁移能力的影响.方法 免疫组织化学法检测EIF4A1在胃癌和癌旁组织中的表达,real time-PCR、Western Blot检测EIF4A1在胃癌细胞(MGC-803)和正常胃黏膜细胞(GES-1)中的表达.在胃癌细胞MGC-803中干扰EIF4A1的表达,采用real time-PCR检测EIF4A1 mRNA表达水平,Western Blot检测EIF4A1蛋白表达水平,采用MTT法和克隆形成实验检测细胞增殖能力,Transwell实验检测细胞迁移能力,West-ern Blot检测E-cadherin,N-cadherin,基质金属蛋白酶2(matrix metallopeptidase 2,MMP2)蛋白表达水平.结果 胃癌组织中EIF4A1的表达量(74.51％)高于癌旁组织(29.03％),胃癌细胞中EIF4A1的表达高于正常胃黏膜细胞,差异均具有统计学意义(P＜0.05).干扰EIF4A1后,实验组(si-EIF4A1)OD值低于对照组(si-NC),实验组平板克隆形成数为33.33±3.51,对照组为56.33±3.51,实验组穿膜细胞数为283.22±43.18,对照组354±88.43,以上结果差异均具有统计学意义(P＜0.05).Western Blot结果显示E-cadherin表达上调,N-cadherin、MMP2表达下调.结论 EIF4A1在胃癌中高表达并与胃癌细胞增殖密切相关,其可能通过EMT相关蛋白和MMP2介导肿瘤细胞迁移.

目的 探讨宫颈癌组织和细胞中WISP2(Wnt-1 inducible secreted protein 2)基因的表达情况并分析WISP2基因是否与HPV感染有关,检测WISP2基因在细胞核中高表达对宫颈癌细胞增殖以及迁移的影响.方法 采用正常宫颈组织37例、CIN(宫颈上皮内瘤变)组织27例及宫颈癌组织35例制作组织芯片,采用免疫组织化学染色(Immu-nohistochemical staining,IHC)对所收集样本中WISP2蛋白质表达情况进行分析.运用HC-2(第二代杂交俘获技术)检测所收集样本HPV(Human papillomavirus)感染情况.构建WISP2过表达质粒,实验组和对照组分别为过表达WISP2组和空质粒组(NC组).Western blot实验和qRT-PCR实验分别检测目的基因在宫颈癌细胞中的表达水平;MTT实验和划痕实验分别对SiHa、HeLa细胞增殖能力以及迁移能力进行测定.结果 WISP2基因在宫颈癌细胞质与核内均有表达,且WISP2基因在CIN 3和宫颈癌组织的细胞质中低表达,在细胞核中高表达(P<0.05);WISP2基因表达与HPV感染具有相关性(P<0.05);WISP2基因在细胞核表达量升高后,细胞增殖、迁移能力均明显增强(P<0.05);WISP2基因细胞核中表达量的升高导致 β-catenin、Vimentin、Slug蛋白质表达量升高,Claudin-1蛋白质表达量降低(P<0.05).结论 WISP2在细胞核表达量增加对促进宫颈癌细胞增殖、迁移和EMT(Epithelial-mesen-chymal transformation)的发生具有重要作用,WISP2基因细胞核中高表达可能是导致宫颈癌发生发展.

目的 研究miR-34a-5p对卡波氏肉瘤相关疱疹病毒(Kaposi's sarcoma-associated herpesvirus,KSHV)感染的神经细胞增殖和迁移能力的影响.方法 选择KSHV感染的SH-SY5Y神经细胞(SK-RG)和SH-SY5Y细胞,进行差异表达的miRNA转录组测序和生物信息学分析,通过qRT-PCR法验证SH-SY5Y、SK-RG中miR-34a-5p表达水平.采用Lipofectamine 2000分别将miR-34a-5p mimics与miR-NC转染至SK-RG细胞,qRT-PCR检测转染效果.采用MTT法及平板克隆实验检测细胞增殖能力.通过Transwell及划痕实验比较两组细胞的迁移能力.采用TargetScan分析miR-34a-5p与候选靶基因c-fos基因3'UTR区的结合位点,采用双荧光素酶报告基因实验检测miR-34a-5p和c-fos 3'UTR区的靶向调控关系以分析其作用机制.采用Western blot在转染miR-34a-5p mimics与miR-NC的细胞中检测靶基因c-fos的表达.结果 转录组测序结果及qRT-PCR验证均提示,SK-RG细胞中miR-34a-5p的水平低于SH-SY5Y细胞(P＜0.05).于SK-RG细胞中过量表达miR-34a-5p后,MTT和平板克隆实验结果表明miR-34a-5p组细胞增殖能力低于miR-NC对照组(P＜0.05).Transwell和细胞划痕实验结果表明,miR-34a-5p组细胞迁移能力低于miR-NC对照组(P＜0.05).双荧光素酶报告基因实验结果提示miR-34a-5p可通过3'UTR区调控c-fos的表达.Western blot结果显示,miR-34a-5p可负向调控c-fos表达.结论 miR-34a-5p可能通过抑制c-fos的表达而抑制KSHV感染的神经细胞的增殖和迁移能力,可能成为治疗KSHV感染相关疾病的小分子miRNA候选药物.

Introduction The aim of this study was to determine whether ZNF582 gene methylation and tissue protein expression can be used as a tool with high sensitivity and specificity for cervical cancer screening. We analyzed the correlation between promoter methylation of ZNF582 gene and cervical cancer and high risk HPV16/18 infection. Methods Tissue samples of normal cervical or chronic cervicitis (n=51), CIN (cervical intraepithelial neoplasia) (n=35), and cervical carcinoma (n=68) were tested for HPV16/18 infection by polymerase chain reaction (PCR). We also detected the methylation status of the ZNF582 gene promoter in the same tissues by methylation-specific PCR (MSP), then analyzed the correlation between ZNF582 promoter methylation and HPV16/18 infection. Immunohistochemistry was used to analyze ZNF582 gene expression in 152 cervical tissues. We detected ZNF582 mRNA expression in cervical tissues (including cancer and non-cancer) by real-time fluorescence quantitative PCR (qPCR). Results Among 93 high-grade cervical lesions (CINII and above) and cervical cancer samples, 57 cases were positive for HPV16/18 infection and 36 cases were negative. ZNF582 gene methylation occurred in 9 out of 51 cases in normal cervical tissues (17.6%), 16 of 35 cases in CIN tissues (45.7%), and 50 of 68 cases in cervical cancer (73.5%). The differences in methylation rate of the three groups were statistically significant (P<0.05). The ZNF582 methylation rate in the positive HPV16/18 infection group was 73.7%, while the negative group was 63.9%. Compared with normal tissues, ZNF582 protein was highly expressed in cervical cancer tissues, but mRNA expression was low. Conclusion While ZNF582 protein is highly expressed in cervical cancer tissues, it was not sufficient for use as a standard for cervical cancer staging. On the other hand, ZNF582 promoter methylation had high specificity and sensitivity in detecting CINII and highly diseased cervical lesions and could be used as a diagnostic marker for cervical cancer of women.