微小RNA-27a在子宫颈癌中的表达变化及作用机制

目的 探讨微小RNA（miR)-27a靶向调控F框/WD-40域蛋白7（FBXW7）对子宫颈癌细胞的增殖、凋亡及侵袭的影响。 方法 30例宫颈癌组织和癌旁组织、30例正常宫颈组织新鲜标本用于实验。Real-time PCR检测宫颈癌、癌旁组织、正常宫颈组织以及宫颈癌细胞（SiHa、Caski、HeLa、HCC94）、子宫颈鳞状上皮永生化细胞H8中miR-27a的表达。应用脂质体转染法将miR-27a抑制剂（inhibitor）及其阴性对照转染至SiHa细胞,CCK-8法、流式细胞术、Transwell法分别检测miR-27a对SiHa细胞增殖活性、细胞周期、凋亡率和侵袭能力的影响。生物学信息法预测miR-27a的靶向基因,双荧光素酶报告基因实验结合Western bloting验证miR-27a对FBXW7的靶向调控作用。 结果 与癌旁组织和正常宫颈组织相比,宫颈癌组织中miR-27a高表达（P<0.05）;与子宫颈鳞状上皮永生化细胞H8相比,宫颈癌细胞SiHa、Caski、HeLa、HCC94中miR-27a高表达（P<0.05）。抑制SiHa细胞中miR-27a的表达,能够明显降低细胞的增殖活性（P<0.05）,提高G0/G1期细胞比例（P<0.05）,降低S期和G2/M期细胞比例（P<0.05）, 提高细胞凋亡率（P<0.05）,抑制细胞的侵袭能力（P<0.05）。生物学信息法预测FBXW7可能是miR-27a的靶向调控基因;双荧光素酶报告基因实验显示,miR-27a可以与FBXW7基因的3’UTR区特异性结合（P<0.05）, 并负调控FBXW7蛋白的表达（P<0.05）。 结论 MiR-27a在宫颈癌的发生发展中起着癌基因的作用,抑制miR-27a表达能够明显抑制宫颈癌细胞的恶性生物学行为,其机制可能与靶向调控FBXW7的表达有关。     

MiR-634 通过mTOR 通路抑制宫颈癌细胞c4-1、caski 增殖并促进其凋亡的研究

目的:探讨MiR-634 通过mTOR 通路抑制宫颈癌细胞c4-1、caski 增殖并促进其凋亡的相关机制。方法:选取30 例宫颈癌组织标本和宫颈癌细胞c4-1、caski 作为研究对象,分析正常组织和宫颈癌组织的MiR-634 和mTOR 表达,MiR-634对宫颈癌细胞mTOR 的表达水平以及对宫颈癌细胞的增殖、迁移和侵袭的影响。结果:宫颈癌组织MiR-634 相对表达水平显著低于正常组织(P<0.05),宫颈癌组织mTOR 相对表达水平显著高于正常组织(P<0.05),宫颈癌组织mTOR 的IS 得分显著高于正常组织(P<0.05);过表达MiR-634 显著降低mTOR 的RNA 和蛋白表达水平(P<0.05),抑制MiR-634 的表达显著提高mTOR 的RNA 和蛋白表达水平(P<0.05);过表达MiR-634 显著降低宫颈癌细胞的增殖、迁移和侵袭(P<0.05),抑制MiR-634的表达显著提高细胞的增殖、迁移和侵袭(P<0.05);抑制mTOR 的表达均显著降低细胞的增殖、迁移和侵袭(P<0.05)。结论:MiR-634 通过抑制mTOR 的表达来抑制宫颈癌细胞的增殖、迁移和侵袭,是宫颈癌基因靶向治疗的潜在靶点。     

miR-155-5p在宫颈癌血清中的表达及其对宫颈癌细胞的影响

 目的：检测miR-155-5p在不同宫颈疾病患者血清中的表达差异,并分析其对宫颈癌细胞增殖、细胞周期和凋亡的影响,探讨miR-155-5p在宫颈癌发生、发展中的可能作用机制。方法：采用SYBR GreenⅠ实时荧光定量PCR法,检测并分析比较miR-155-5p在不同宫颈疾病患者血清中的表达差异。利用miR-155-5p mimic或inhibitor提高或降低宫颈癌细胞中miR-155-5p的表达。CCK-8法和流式细胞术检测宫颈癌细胞的增殖、细胞周期和凋亡。结果：宫颈癌组血清中miR-155-5p的表达高于宫颈炎组和健康对照组（P<0.05）,宫颈上皮内瘤样病变组和宫颈癌组血清中miR-155-5p的表达差异无统计学意义（P>0.05）。与空白组、脂质体组和阴性对照组相比,转染100 nmol/L和200 nmol/L miR-155-5p mimic的SiHa细胞中,S期细胞比例升高,凋亡细胞比例降低（P<005）。转染100 nmol/L和200 nmol/L miR-155-5p inhibitor的SiHa细胞中,G2/M期细胞比例明显增多（P<005）。结论：(1)宫颈癌患者血清中miR-155-5p表达较健康对照人群上调,可能作为宫颈癌早期诊断的肿瘤分子标志物。(2)miR-155-5p对宫颈癌HeLa细胞增殖、细胞周期和凋亡无明显影响。(3)miR-155-5p可促进宫颈癌SiHa细胞进入S期,并抑制SiHa细胞凋亡,提示miR-155-5p可能在宫颈鳞癌发生、发展中起作用。     

miR-96 enhances the proliferation of cervical cancer cells by targeting FOXO1

MiRNAs affect various biological pathways associated with the development, progression, clinical outcome and treatment response improvement in cervical cancer. This study was performed to evaluate the effects of miRNA 96 on cervical cancer and to clarify the mechanism. Vivo and vitro experiments were conducted in our trial. MiR-96 is upregulated in cervical cancer cell lines and cervical cancer tissues and is correlated with clinical features in cervical cancer patients. Overexpression of miR-96 enhances proliferation of cervical cancer cells, while inhibiting miR-96 reduces the proliferation of cervical cancer cells. Inhibition of miR-96 significantly decreased the percentage of cells in the S phase and increased the percentage of cells in G1/G0 peak in both SiHa and CaSki cells compared with NC cells and decreased the expressions of p21, p27 and cyclin D1. FOXO1 3’-UTR was sub cloned into a luciferase reporter vector and the putative miR-96 binding site in the FOXO1 3’-UTR was mutated. Treated with miR-96 inhibitor consistently enhanced the luciferase activity of the FOXO1 3’-UTR luciferase reporter plasmids in both SiHa and CaSki cells, whereas mutations in the miR-96-binding site abolished the effect. Vivo experiment also support these results. Therefore, inhibition of miR-96 might suppress growth, proliferation of CC cells and promote apoptosis of CC cells both in vitro and in vivo.     

lncRNA PSMA3-AS1 靶向 miR-3619-5p 调控宫颈癌 SiHa 细胞增殖、迁移及侵袭的分子机制研究

目的 探讨 lncRNA PSMA3-AS1 调控宫颈癌细胞增殖、 迁移及侵袭的分子机制。 方法 收集 2020 年 3 月至 2021 年 6 月西安医学院第二附属医院收治的 42 例宫颈癌患者的癌组织及其癌旁组织标本, 采用 qRT-PCR 法检测宫颈癌组织、 癌旁组织、 人宫颈上皮永生化细胞 H8、 与人宫颈癌细胞系 SiHa、 HeLa、 Caski 中 lncRNA PSMA3-AS1、 miR-3619-5p 的表达量; 以 SiHa 细胞为研究对象, 分组为: si-NC 组、 si-lncRNA PSMA3-AS1 组、 miR-NC 组、 miR-3619-5p 组、 anti-miR-NC + si-lncRNA PSMA3-AS1 组、 anti-miR-3619-5p + si-lncRNA PSMA3-AS1 组; MTT 法与 Transwells 实验分别检测每组 SiHa 细胞的增殖活力、 迁移及侵袭能力; 双 荧光素酶报告实验检测 miR-3619-5p 过表达对野生型载体 lncRNA PSMA3-AS1-WT、 突变型载体 lncRNA PSMA3-AS1-MUT 荧光素酶活性的影响; Western 印迹检测 MMP2、 MMP9 蛋白表达量。 结果 宫颈癌组织中 lncRNA PSMA3-AS1 的表达量比癌旁组织增加 ( P < 0. 01), miR-3619-5p 的表达量比癌旁组织减少 ( P < 0. 01); 与 H8 细胞比较, SiHa、 HeLa、 Caski 细胞中 lncRNA PSMA3-AS1 的表达量升高 (P< 0. 01), miR-3619-5p 的表达量降低 (P< 0. 01); 与 si-NC 组比较, si-lncRNA PSMA3-AS1 组细胞活力和 MMP2、 MMP9 蛋 白水平降低 (P< 0. 05), 迁移及侵袭细胞数减少 (P< 0. 05); 与 miR-NC 组比较, miR-3619-5p 组细胞活力 和 MMP2、 MMP9 蛋白水平降低 (P< 0. 01), 迁移及侵袭细胞数减少 (P< 0. 05); miR-3619-5p 过表达可抑 制 lncRNA PSMA3-AS1-WT 的荧光素酶活性 (P< 0. 01), 而未能影响 lncRNA PSMA3-AS1-MUT 的荧光素酶活 性; 与 anti-miR-NC + si-lncRNA PSMA3-AS1 组比较, anti-miR-3619-5p + si-lncRNA PSMA3-AS1 组细胞活力和 MMP2、 MMP9 蛋白水平升高 (P< 0. 01), 迁移及侵袭细胞数增多 (P< 0. 01)。 结论 干扰 lncRNA PSMA3- AS1 表达可通过促进 miR-3619-5p 而降低宫颈癌细胞增殖、 迁移及侵袭能力。     

微小RNA-513c-5p在宫颈癌中的表达及靶向组蛋白去乙酰化酶1调节宫颈癌细胞迁移和侵袭的机制

目的 探讨微小RNA（miR)-513c-5p在宫颈癌中的表达及靶向组蛋白去乙酰化酶1（HDAC1）调节宫颈癌细胞迁移和侵袭的机制。方法 临床收集宫颈癌患者86例,通过Real-time PCR检测肿瘤组织和癌旁组织中miR-513c-5p水平,分析其与宫颈癌病理特征的关系。通过双荧光素酶报告验证miR-513c-5p靶向HDAC1。将宫颈癌HeLa细胞系分为4组：对照组、类似物(mimic)组、mimic+HDAC1组和HDAC1组。通过质粒转染技术过表达miR-513c-5p和(或)HDAC1。Real-time PCR和Western blotting分别用于检测RNA或蛋白的表达水平。分别通过CCK-8法、细胞划痕实验和Transwell实验检测各组的细胞生长、迁移和侵袭能力。 结果 宫颈癌组织中miR-513c-5p水平显著低于癌旁组织。低水平的miR-513c-5p与更高的局部侵袭、淋巴转移和远端转移有关（P<0.05）。miR-513-5p靶向抑制HDAC1表达。过表达miR-513c-5p显著抑制宫颈癌细胞生长、迁移和侵袭（P<0.05）。过表达HDAC1促进细胞生长、迁移和侵袭（P<0.05）,并且可以逆转miR-513c-5p的抑制作用（P<0.05）。 结论 低水平的miR-513c-5p可能与宫颈癌转移有关,并且miR-513c-5p可通过靶向抑制HDAC1蛋白的表达抑制宫颈癌HeLa细胞生长、迁移和侵袭。     

MST1过表达抑制宫颈癌细胞系SiHa的增殖、迁移和侵袭

目的探讨哺乳动物不育系20样激酶1(MST1)对子宫颈癌SiHa细胞增殖、迁移和侵袭能力的影响。方法Western blot检测正常宫颈上皮细胞H8与宫颈癌细胞SiHa中MST1的表达;构建p J3H-HA-MST1质粒并转染SiHa细胞系,Western blot检测MST1、Ki-67和MMP9的蛋白表达;MTS、划痕实验和Transwell分别检测细胞增殖、迁移和侵袭能力。结果 SiHa细胞的MST1蛋白表达水平明显低于H8细胞(P0.05);SiHa细胞转染MST1质粒后,MST1蛋白表达明显升高,而Ki-67和MMP9蛋白表达水平显著下降(P0.05),SiHa细胞的增殖被明显抑制(P0.05),同时细胞的迁移和侵袭能力也显著降低(P0.01)。结论 MST1过表达可以抑制宫颈癌细胞SiHa的增殖、迁移和侵袭能力。     

MiR-211 inhibits cell proliferation and invasion of gastric cancer by down-regulating SOX4

Introduction: Previous studies have shown that the dysregulation of miRNAs are frequently associated with cancer progression. Deregulation of miR-211 has been observed in various types of human cancers. However, its biological function in gastric cancer (GC) is still unknown. Methods: The expression of miR-211 in GC was detected by using quantitative real-time PCR (qRT-PCR). The miR-211 mimics and inhibitor were designed and transfected into BGC-823 cells. Then, we explore the probable biological function of miR-211 in gastric cancer cell proliferation and invasion in vitro. A luciferase reporter assay and western blot were performed to confirm the target gene of miR-211. Results: MiR-211 was significantly down-regulated in GC. Over-expression of miR-211 inhibited gastric cancer cell proliferation and invasion in vitro, conversely, down-regulated expression of miR-211 promoted gastric cancer cell proliferation and invasion. In addition, the sex-determining region Y-related high mobility group box 4 (SOX4) is identified as a target of miR-211 in GC cells, and SOX4 expression levels was inversely correlated with miR-211. Furthermore, knockdown of Sox4 inhibited the proliferation and invasion in GC cells. Conclusion: miR-211 could inhibit GC cell proliferation and invasion partially by down-regulating SOX4. MiR-211 might be a potential therapeutic target for GC treatment in the future.     

miRNA-451在膀胱癌细胞中的表达及对其生物学功能的影响

目的：检测微小RNA(microRNA,miRNA)在膀胱癌细胞中的表达及探讨其对膀胱癌细胞迁移、侵袭、黏附及增殖能力的影响。方法：采用实时荧光定量PCR(quantitative Real-time PCR,qPCR)检测miR-451在不同转移潜能膀胱癌细胞株T24、5637、J82中的表达;使用Lipo-2000脂质体将miR-451拟似物(miR-451 mimics)转染入5637细胞,qPCR验证其转染效率,采用划痕愈伤实验、Transwell实验、细胞黏附及MTT增殖实验分别检测细胞二维迁移、侵袭、黏附及增殖能力的变化。结果：miR-451在T24、5637及J82中的相对表达量分别为0.06±0.001、0.13±0.024、1(将J82细胞中其表达量标准化为1),差异显著,具有统计学意义(P<0.01);miR-451过表达后,5637细胞的迁移、侵袭、黏附能力及增殖率显著降低,具有统计学意义(P<0.05)。结论：miR-451在不同膀胱癌细胞中差异表达,其表达异常可影响癌细胞生物学功能,这为膀胱癌靶向分子治疗提供了新的切入点。     

MicroRNA-125b在胃癌组织中的高表达并促进胃癌细胞系HGC-27和MGC-803的增殖

 目的 探讨microRNA-125b (miR-125b)基因在胃癌患者组织中的表达改变情况,及其对胃癌细胞系增殖和凋亡的影响。方法 使用real-time PCR方法检测miR-125b在40例临床诊断为胃癌患者的癌组织与癌旁对照组织中的表达情况。随后使用miR-125b mimic转染胃癌细胞系HGC-27和MGC-803,确认过表达成功后,分别使用CCK-8试剂盒和流式细胞仪检测过表达miR-125b对细胞增殖和凋亡的影响。结果 证实在胃癌患者组织中miR-125b的表达水平显著高于癌旁对照组（P<0.01）。在胃癌细胞系HGC-27和MGC-803中过表达miR-125b后,细胞增殖明显增加：转染72h,HGC-27（scramble组：1.632±0.09,mimic组：2.473±0.08）,MGC-803（scramble组：1.603±0.05,mimic组：2.554±0.07））,同时细胞凋亡也受到抑制。结论 miR-125b可能作为癌基因在胃癌中发挥作用,并对细胞增殖和凋亡具有显著影响。     

miR-1246增强宫颈癌细胞辐射敏感性的分子机制研究

目的:探讨微小RNA-1246(miR-1246)过表达增强宫颈癌细胞放疗敏感性的分子机制。方法:运用脂质体2000将miR-1246模拟物(miR-1246 mimic)转染4种宫颈癌细胞系He La、Ca Ski、C33A和Si Ha,以阴性对照模拟物(NC-mimic)为阴性对照。Real-time PCR检测宫颈癌组织、正常组织、子宫内膜上皮细胞系ESC和4株宫颈癌细胞中miR-1246的表达水平;转染的细胞经电离辐射照射后运用MTT法和Transwell法分别测定细胞活力和细胞迁移能力;运用免疫荧光法检测γH2AX表达水平;Western blot检测细胞中γH2AX、ATM、p-ATM和p-p53的蛋白水平。结果:miR-1246在正常组织和ESC细胞中高表达,而在4株宫颈癌细胞和宫颈癌组织中低表达;转染miR-1246 mimic后miR-1246表达水平显著高于NC-mimic组细胞(P 0. 05)。相同条件下,辐照后miR-1246过表达组宫颈癌细胞活力显著低于NC-mimic组,细胞迁移率明显降低(P 0. 05)。免疫荧光结果显示,miR-1246过表达显著增强电离辐射诱导的γH2AX激活(P 0. 05);Western blot结果显示,与NC-mimic组比较,miR-1246过表达显著促进电离辐射诱导γH2AX蛋白的表达,减低p-ATM和p-p53的蛋白水平(P 0. 05)。结论:miR-1246在正常组织和子宫内膜上皮细胞中高表达,而在宫颈癌组织和宫颈癌细胞系中低表达;miR-1246过表达抑制宫颈癌细胞活力和迁移能力,并可能通过阻断ATM通路、抑制DNA损伤修复而增强宫颈癌细胞的辐射敏感性。     

STAT3短发夹RNA表达载体的构建及对SiHa细胞增殖和凋亡的作用

目的:探讨信号转导子与转录激活子3短发夹RNA(shRNA)真核表达载体对宫颈癌SiHa细胞增殖和凋亡的作用.方法:根据siRNA设计原则, 结合pSilencer2.1-U6-neo质粒特点, 针对STAT3基因设计并合成两条寡聚DNA片段, 退火后克隆入pSilencer2.1-U6-neo质粒, 脂质体法将重组质粒转染人宫颈癌SiHa细胞.RT-PCR和Western blot检测SiHa细胞STAT3基因mRNA和蛋白表达水平;MTT法和流式细胞术(FCM)检测细胞的增殖和凋亡.结果:成功构建了STAT3基因shRNA真核表达载体, 转染人宫颈癌SiHa细胞.细胞STAT3蛋白及mRNA表达下降;同时SiHa细胞增殖能力下降, 细胞凋亡增加.结论:构建的STAT3基因shRNA真核表达载体能够通过下调STAT3基因表达, 抑制宫颈癌SiHa细胞的增殖能力, 诱导细胞凋亡.     

miR-375 is down-regulated in squamous cervical cancer and inhibits cell migration and invasion via targeting transcription factor SP1

Pelvic lymph node metastases are regarded as the most important risk factor and a predictor of poor prognosis for patients with cervical cancer. Exploration of metastasis-related molecules is helpful toward improving the prognosis in cervical cancer. To identify the role of miR-375 in metastasis and progression of cervical cancer, we examined the expression of miR-375 in 170 cervical cancer tissues and 68 normal cervical tissues, using stem-loop quantitative PCR, and found that the expression of miR-375 in cervical cancer tissues was significantly decreased by 4.45-fold, compared with 68 normal tissues. A significant correlation existed between miR-375 expression and clinicopathologic parameters, including lymph node metastasis of cervical cancer. Overexpressed miR-375 suppressed cell proliferation, blocked G1-to-S cell-cycle transition, and inhibited cell migration and invasion in human cervical SiHa and CaSki cells. SP1, a potential target gene of miR-375, was inversely correlated with miR-375 expression in cervical cancer tissues. Moreover, SP1 was negatively regulated by miR-375, and knockdown of SP1 by siRNA inhibited cell malignant behaviors. Thus, our findings suggest that down-regulated miR-375 promotes cell malignant behaviors via the target gene SP1 and may consequently contribute to the progression of cervical cancer.     

MiR-150-5p inhibits the proliferation and promoted apoptosis of pancreatic cancer cells北大核心CSCD

Objective: To investigate the role of miR-150-5p in cell proliferation and apoptosis in human pancreatic cancer cell lines. Methods: The expression of miR-150-5p in pancreatic cancer was detected by real time qPCR analysis in 11 pairs of pancreatic cancer tissue and matched adjacent normal tissue samples and in 4 pancreatic cancer cell lines. PANC-1, MIA PaCa-2, BxPC-3 and AsPC-1 cells were transfected with chemically synthesized MiR-150-5p mimics, and CCK-8 assays was then performed to assess cellular functions. To fully understand the mechanisms by which miR-150-5p exerted its function, cell cycle analysis was performed on MIA PaCa-2 and PANC-1 cells 48 hours after transfection, by incubating with propidium iodide (PI) and subsequently analyzed by fluorescence-activated cell sorting (FACS). Apoptosis assay was performed on MIA PaCa-2 and PANC-1 cell lines 24 hours after transfection using the Annexin V-FITC Apoptosis Detection Kit I (BD Biosciences) and analyzed by FACS. Results: The expression of miR-150-5p was consistently lower in the pancreatic cancer tissues than in normal tissues,and the miR-150-5p was also down-regulated in pancreatic cancer cell lines (P < 0. 05). MiR-150-5p mimics transfection significantly raised the expression level of miR-150-5p mRNA in PANC-1 and MIA PaCa-2 (P <0. 01). The CCK-8 proliferation assay showed that cell growth was reduced in 4 pancreatic cancer cell lines (AsPC-1, BxPC-3, MIA PaCa-2, PANC-1) of miR-150-5p transfected cells compared with NC-transfected cells. The inhibition rates were 50. 7%,48. 6%,30. 8% and 42. 3%, respectively (P <0. 01). The apoptotic rate was increased in cells transfected with miR-150-5p mimics (P < 0. 01). The cell cycle analysis in MIA PaCa-2 indicated that miR-150-5p treatment induced cell cycle arrest in G1 phase with a significant increase in the percentage of cells in G, phase (P < 0. 01), and a reduction of the S-phase cell population in MIA PaCa-2 and PANC-1 (P<0.01). Conclusions: MiR-150-5p is down-regulated in pancreatic cancer. Overexpression of miR-150-5p inhibits cell proliferation, blocked the cell cycle, but promotes cell apoptosis in pancreatic cancer cells.     

miR-101 suppresses tumor proliferation and migration,and induces apoptosis by targeting EZH2 in esophageal cancer cells

Aim: To investigate the role of miR-101 in the regulation of tumor proliferation, invasion, apoptosis and to its target gene in human ESCC. Methods: The expression level of miR-101 in Eca109 cell line was determined by real-time polymerase chain reaction (PCR). After transfected with miR-101 mimics and inhibitor, proliferation, migration and apoptosis in ESCC cell line (Eca109) were detected by MTT, cell wound healing assay and flow cytometry, respectively. The expression of EZH2 in Eca109 cell was examined by immunohistochemical staining. Results: We found that miR-101 was significantly down-regulated in ESCC cell than in matched normal esophageal epithelium cell. The expression level of miR-101 was inversely correlated to EZH2 protein expression in ESCC cell. In Eca109 cells, over-expression of miR-101 significantly inhibited the migration and invasion of ESCC cells, and promotes cell apoptosis. Conclusions: These findings suggest that decreased expression of miR-101 might promote metastasis of human ESCC by inducing accumulation of EZH2 protein.     

miR-218 调控SNX4 蛋白对乳腺癌细胞增殖和侵袭的影响

目的：探讨miR-218调控SNX4蛋白对乳腺癌细胞增殖和侵袭能力的影响。 方法：qPCR检测乳腺癌组织和乳腺癌细胞株中miR-218的表达情况;分析miR-218的表达和乳腺癌临床病理参数之间的关系;双荧光素酶实验检测miR-218和SNX4之间的关系;过表达miR-218后MTT实验和侵袭实验检测乳腺癌细胞增殖和侵袭能力变化;过表达SNX4后MTT实验和侵袭实验检测SNX4对乳腺癌细胞增殖和侵袭能力的恢复水平;裸鼠体外成瘤实验检测miR-218对乳腺癌细胞株成瘤能力的影响。结果：miR-218在乳腺癌组织和MCF-7细胞株中表达水平较高,miR-218的表达与乳腺癌的病理分期相关以及淋巴结转移情况有关, SNX4可能是miR-218下游的作用靶点;过表达miR-21可以抑制乳腺癌细胞的增殖侵袭能力,过表达SNX4后可以逆转miR-218对乳腺癌细胞的抑制作用,过表达miR-218后可以抑制乳腺癌细胞株在裸鼠体内的成瘤能力。结论：miR-218在乳腺癌中表达上调,同时miR-218可以调控SNX4的表达而影响乳腺癌细胞增殖和侵袭能力。     

微小RNA-98-5p靶向调控核糖核苷酸还原酶小亚基M2调控宫颈癌细胞顺铂的耐药性

目的 探讨微小RNA(miR)-98-5p对顺铂(DDP)耐药宫颈癌细胞顺铂敏感性的调控及其机制.方法 用脂质体法将 DDP+miR-NC 组(转染 miR-NC)、DDP+miR-98-5p 组(转染 miR-98-5p mimics)、DDP+si-NC 组(转染si-NC)、DDP+si-核糖核苷酸还原酶小亚基M...     

miR-30c抑制宫颈癌细胞恶性表型的分子机制研究

目的:探讨微小RNA(miR)-30c过表达抑制宫颈癌细胞恶性表型的分子机制。方法:运用Lipofectamine 2000转染法将pGenesil-1-miR-30 c质粒转染宫颈癌细胞系C33A、HeLa、SiHa和CaSki,以转染p-Geresil-1的细胞为阴性对照;运用TaqMan real-time PCR检测各组细胞中miR-30c的表达水平;采用MTT法、集落形成实验、Transwell法、Annexin V-FITC及流式细胞术分别测定细胞活力抑制率、集落形成能力、迁移率及凋亡率;Western blot检测Bax、Bcl-2、基质金属蛋白酶(MMP)-9、MMP-13和金属蛋白酶组织抑制物1(TIMP-1)的蛋白表达水平。结果:转染p Genesil-1-miR-30c质粒的宫颈癌细胞系中miR-30c表达水平均显著高于阴性对照组(P0.01);过表达miR-30 c的宫颈癌细胞活力抑制率显著高于阴性对照组(P0.05),细胞集落形成率和迁移率显著低于阴性对照组(P0.05);流式细胞术检测结果显示,miR-30c过表达的宫颈癌细胞凋亡率显著高于对照组(P0.05);Western blot结果显示miR-30 c过表达促进Bax和TIMP-1蛋白的表达,而抑制Bcl-2和MMP-13蛋白的表达(P0.05或P0.01)。结论 :miR-30 c过表达抑制宫颈癌细胞活力和迁移,诱导宫颈癌细胞凋亡,其机制与激活细胞凋亡通路及抑制MMP-13表达有关。     

miR-145过表达对宫颈癌细胞辐射敏感性的影响

目的:探讨微小RNA(miR)-145过表达增强宫颈癌细胞放疗敏感性的分子机制。方法:运用Lipofectamine 2000将miR-145-mimic转染4株宫颈癌细胞系He La、Ca Ski、C33A和Si Ha,以NC-mimic为阴性对照,并以real-time PCR检测子宫内膜基质细胞(ESC)和4株宫颈癌细胞中miR-145的表达水平;转染的细胞经电离辐射照射后于不同时点运用MTT法和Annexin V-FITC/PI染色法联合流式细胞术分别测定细胞活力和细胞凋亡率;运用免疫荧光检测组蛋白γH2AX的表达水平;Western blot检测细胞中螺旋酶样转录因子(HLTF)的蛋白表达水平。结果:miR-145在ESC中高表达,而在4株宫颈癌细胞中均呈低表达,转染miR-145-mimic后4株宫项癌细胞中miR-145的表达水平显著高于NC-mimic组细胞(P0.05)。相同条件下,辐照后miR-145过表达的宫颈癌细胞的活力显著低于NC-mimic组细胞,72 h的细胞凋亡率明显增加(P0.05)。免疫荧光观察结果显示miR-145过表达显著促进电离辐射诱导的γH2AX激活;Western blot结果显示,miR-145过表达显著抑制HLTF的表达,与NC-mimic比较差异有统计学意义(P0.05)。结论:miR-145在正常子宫内膜上皮细胞中高表达,而在宫颈癌细胞系中低表达;miR-145过表达可显著抑制宫颈癌细胞的活力,促进电离辐射诱导的细胞凋亡。miR-145过表达增强宫颈癌细胞的辐射敏感性,其机制可能与下调HLTF表达、抑制DNA损伤修复、促进细胞凋亡有关。