Study on the expression of miRNA-126 in the renal cell carcinoma and the effects on biological characteristics of cell line 786-O
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摘要: 目的:研究MicroRNA-126(miRNA-126)在肾透明细胞癌(ccRCC)中的表达及其对细胞株786-O生物学行为的机制。方法:通过Real-time PCR的方法检测30例ccRCC及癌旁正常肾组织中miRNA-126-5p的表达情况。运用RT-qPCR的方法检测肾癌细胞株786-O和人肾小管上皮细胞株HK-2中miRNA-126-5p的表达;用Lipofectamine RNAiMAX将hsa-miRNA-126-5p转染786-O细胞株,用MTT、Transwell侵袭实验、Annexin V/PI双染法观察转染后细胞在增殖、侵袭力和凋亡方面的变化。结果:miRNA-126-5p在肾癌组织中的表达明显降低,在不同病理分级的样本中,miRNA-126-5p在高分化组中表达量为36.63±5.38,中、低分化组为19.96±7.28,差异无统计学意义(P＞0.05)。在不同临床分期的样本中,Ⅰ期miRNA-126-5p表达量为37.05±6.38,Ⅱ~Ⅲ期的表达量为24.03±6.15,不同分期之间差异无统计学意义(P＞0.05)。786-O细胞株转染后,检测miRNA-126-5p在实验组细胞的表达为42.86±7.01,显著高于阴性对照组27.46±8.43(P＜0.05)。MTT法检测786-O转染后48h增殖情况,实验组的细胞相对存活率为80.82±0.47,阴性对照组98.87±0.27,差异有统计学意义(P＜0.01)。Transwell侵袭实验显示,实验组miRNA-126-5p表达明显低于阴性对照组[(33.89±2.80) vs.(53.67±2.23),P＜0.05]。Annexin V/PI双染法检测细胞凋亡,实验组的细胞凋亡率明显高于阴性对照组[(22.56±1.13) vs.(7.20±0.26),P＜0.05]。结论:miRNA-126-5p在ccRCC组织中的表达量明显低于癌旁组织中的表达,其表达与肿瘤分级和临床分期无明显相关性。hsa-miRNA-126-5p转染细胞株786-O后,miRNA-126-5p的表达明显上调,并能够使786-O细胞增殖受到抑制,侵袭力降低,凋亡率增加。
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关键词:
- 肾透明细胞癌 /
- miRNA-126 /
- Real-timePCR /
- 肾癌细胞株786-O /
- 细胞增殖 /
- 细胞侵袭力 /
- 细胞凋亡
Abstract: Objective: To investigate the expression of miRNA-126 in clear cell renal cell carcinoma (ccRCC) tissues and to study the effects of miRNA-126 in 786-O cells.Method: For real-time qPCR, the expressions of miRNA-126 were performed in 30 cases of ccRCC and normal adjacent tissues.The expressions of miRNA-126-5p were detected by RT-qPCR in 786-O cells and HK-2 cells.The 786-O cells were transfected by hsa-miRNA-126-5p using Lipofectamine RNAiMAX to up-regulate the expression of miRNA-126-5p.To study the changes of cell proliferation, invasion ability and apoptosis by MTT, transwell invasion test and flow cytometry respectively.Result: The expression of miRNA-126-5p in ccRCC was lower (P＜0.05). Grouped by the pathological class, the expression of miRNA-126-5p was (36.63±5.38) in pathological class Ⅰ, and (19.96±7.28) in pathological classⅡ-Ⅲ, which had no significant correlations (P＞0.05). In terms of clinical stage, the expression of miRNA-126-5p was (37.05±6.38) in clinical stageⅠ, and (24.03±6.15) in clinical stageⅡ-Ⅲ, which had no significant difference (P＞0.05). After transfection, the expression of miRNA-126-5p in treatment group cells (42.86±7.01) was significantly higher than that in control group (27.46±8.43) (P＜0.05). After 48 hours transfection, the 786-O cell survival rate in treatment group (80.82±0.47) was lower than that in control group (98.87±0.27), has significant difference (P＜0.01). Transwell invasion experiment outcomes showed that the cells numbers in treatment group (33.89±2.80) was significantly lower than that in control group (53.67±2.23) (P＜0.05). The cell apoptosis assay showed that the cell apoptosis rate in treatment group was (22.56±1.13), while the cell apoptosis rate in control group was (7.20±0.26), which had significant difference compared with control group (P＜0.05). Conclusion: The expression of miRNA-126-5p in ccRCC was lower than that in its adjacent normal tissues.No significant correlations were observed between miRNA-126-5p expression and clinical stage, pathological class from patients with ccRCC.The expression of miRNA-126-5p in 786-O cells which were successfully transfected by hsa-miRNA-126-5p was significantly up-regulated.After up-regulating miRNA-126-5p, the cells proliferation, invasion ability were significantly suppressed. -
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