Shi Yu-Rong, Liu Jian, He Wei, Yang Yin
Department of Biochemistry and Molecular Biology, Bengbu Medical College,Bengbu 233000,China.
Research Center of Clinical Diagnostics Laboratory, Bengbu Medical College,Bengbu 233000,China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2016 Sep;47(5):697-702.
To investigate the expression and clinical significance of microRNA-218(miR-218)in human cervical cancer and the effects of-218 on proliferation, cell apoptosis and invasion of HeLa cells.
QRT-PCR was used to detect the expression of-218 in 23 cases of normal cervical tissues and 114 cases of cervical cancer, and the relationship between the expression and the clinicopathological features was analyzed; HeLa cells were devided into three groups: non transfection (control group), transfected with empty liposomes negative control goup (NC group), transfected with miR-218 mimic (miR-218M group). The cell growth inhibiting ratio of HeLa cells was assessed by MTT assay. Fluorescence activated cell sorting was used to measure cell apoptosis. Changes of cell migration ability were detected by wound healing test and Transwell assay. QRT-PCR and Western blot were used to detect the expression of Bcl-2, Bax, NF-κB and E-cadherin, respectively.
The expression of-218 in cervical cancer was down regulated, and there were significant differences in the different pathological types, stages, lymph node metastasis and interstitial infiltration in cervical cancer tissues ( <0.01); After being transfected with miR-218 mimic, the proliferation of HeLa cells was significantly inhibited. The ability of invasion was decreased. QRT-PCR and Western blot showed that after being transfected with miR-218 mimic, the expression levels of -2 mRNA and protein were down-regulated and mRNA and protein expression levels were increased, E-cadherin mRNA and protein expression were up-regulated, but mRNA and protein expression were down-regulated.
he low-expression of miR-218 is correlated with the poor clinicopathological features in human cervical cancer. MiR-218 overexpression reduces cancer cell proliferation and induces apoptosis and inhibits cell migration, suggesting that miR-218 may play a key role in the progression of human cervical cancer.
探讨微小RNA-218（miR-218）在人宫颈癌中的表达及临床意义，以及miR-218对HeLa细胞增殖、细胞凋亡和侵袭的影响。
采用实时定量聚合酶链反应（QRT-PCR）检测23例正常宫颈组织和114例宫颈癌组织中miR-218的表达，并分析其表达与临床病理特征的关系；将HeLa细胞分为三组：未转染组（对照组）、转染空脂质体阴性对照组（NC组）、转染miR-218模拟物组（miR-218M组）。采用MTT法评估HeLa细胞的生长抑制率。采用荧光激活细胞分选术检测细胞凋亡。通过伤口愈合试验和Transwell试验检测细胞迁移能力的变化。分别采用QRT-PCR和蛋白质免疫印迹法检测Bcl-2、Bax、核因子κB（NF-κB）和E-钙黏蛋白的表达。
宫颈癌组织中miR-218表达下调，在不同病理类型、分期、淋巴结转移及间质浸润的宫颈癌组织中差异有统计学意义（P<0.01）；转染miR-218模拟物后，HeLa细胞增殖明显受到抑制，侵袭能力下降。QRT-PCR和蛋白质免疫印迹法显示，转染miR-218模拟物后，Bcl-2 mRNA和蛋白表达下调，Bax mRNA和蛋白表达上调，E-钙黏蛋白mRNA和蛋白表达上调，但NF-κB mRNA和蛋白表达下调。
miR-218低表达与人宫颈癌不良临床病理特征相关。miR-218过表达可降低癌细胞增殖、诱导凋亡并抑制细胞迁移，提示miR-218可能在人宫颈癌进展中起关键作用。
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