microRNA-545抑制宫颈癌Hela细胞增殖和凋亡的影响

【摘 要】目的 探讨microRNA-545（miR-545）在宫颈癌组织及细胞系中的表达情况，并探讨其对宫颈癌Hela细胞生物学行为的影响。方法 采用实时定量PCR（qPCR）检测52例宫颈癌组织、30例宫颈上皮内瘤样变（CIN）组织和25例正常宫颈组织石蜡包埋组织中的miR-545表达水平，分析miR-545表达与宫颈癌临床病理特征的关系，并检测其在宫颈癌细胞株Hela、Siha、CaSki、C33a及宫颈正常上皮细胞中的表达情况。采用miR-545的特异性过表达载体pcDNA6.2／GW／EmGFP-miR-545转染miR-545表达水平最低的宫颈癌细胞（过表达组），同时设pcDNA6.2/GW/EmGFP-miR空载体组和空白对照组。分析转染后Hela细胞的增殖水平（四甲基偶氮唑盐法）、凋亡情况（流式细胞仪Annexin V／PI双染法）及细胞周期（流式细胞仪PI单染法）。结果 宫颈癌组织中miR-545的相对表达量为0.245±0.051，低于CIN组织的0.761±0.073和正常宫颈组织的1.027±0.024，差异有统计学意义（P＜0.05），且宫颈癌组织中miR-545水平与临床分期、分化程度及淋巴结转移有关（P＜0.05）；与宫颈正常上皮细胞相比，宫颈癌细胞的miR-545水平均降低，由高到低依次为CaSki、C33a、Siha和Hela细胞；与空载体组和空白对照组相比，转染过表达载体细胞的增殖抑制率、凋亡率及G0／G1期细胞比例均升高，而S、G2／M期细胞比例均降低，差异有统计学意义（P＜0.05）。结论 miR-545在宫颈癌组织和细胞中均为低表达，上调其水平可达到抑制增殖、诱导凋亡和阻滞细胞周期的作用，在宫颈癌防治中具有一定价值。

Effect of microRNA-545 on the proliferation and apoptosis of cervical cancer Hela cells

【Abstract】Objective To investigate the expression of microRNA-545（miR-545）in cervical cancer tissues and cell lines as well as its effect on the biological behavior of cervical cancer Hela cells. Methods The real time quantitative PCR（qPCR）was used to detect the level of miR-545 in 52 cases of cervical cancer，30 cases of cervical intraepithelial neoplasia（CIN）and 25 cases of normal cervical tissues. The relationship between miR-545 expression and clinical pathological parameters of cervical cancer was analyzed. The expression of miR 545 in cervical cancer cell lines（Siha，Hela，CaSki，C33a）and normal epithelial cells was also detected. The miR 545 expression vector pcDNA6.2／GW／EmGFP-miR-545 was transfected into cervical cancer cells with the lowest level of miR-545(over expression group). Meanwhile, the pcDNA6.2/GM/EmGFP-miR empty control group and black control group were set up. The proliferation level（methylthiazolyl tetrazolium），apoptosis（flow cytometry with Annexin V／PI double staining） and cell cycle（flow cytometry with PI staining）of Hela cells were analyzed. Results The relative level of miR-545 was 0.245±0.051 in cervical cancer tissues，lower than 0.761±0.073 in CIN tissues and 1.027±0.024 in normal cervical tissue with statistical significance（P＜0.05）. The miR-545 level in cervical cancer tissues was associated with clinical stage，differentiation degree and lymph node metastasis（P＜0.05）. Compared with normal cervical cells，the miR-545 levels of cervical cancer cells were decreased，which were followed by CaSki，C33a，Siha and Hela cells. Compared with empty control group and blank control group，the cell proliferation inhibition rate，apoptosis rate and the percentage of G0／G1 phase cells were increased but the proportion of S and G2／M phase cells were decreased in over-expression group with statistical difference（P＜0.05）. Conclusion miR-545 is low expressed in cervical cancer tissues and cells，and up-regulation of miR-545 exhibited the effect of inhibiting proliferation,inducing apoptosis and arresting cell cycle，which has certain value in the prevention and treatment of cervical cancer.