MicroRNA-93在宫颈癌中的表达及其对细胞生物学行为的影响

Characterization of miR-93 expression and investigation of its biological function in cervical cancer

摘要:

目的  探讨microRNA-93（miR-93）在宫颈癌组织中的表达及其对宫颈癌细胞生物学行为的影响。方法  收集2014年1月-2014年7月中南大学湘雅医院有完整病历资料的41例宫颈癌患者组织标本及对应癌旁组织，实时荧光定量-聚合酶链反应（qRT-PCR）检测宫颈癌组织中miR-93表达水平；采用脂质体转染法将miR-93模拟片段（mimic）转染入宫颈癌Hela细胞，恢复细胞内miR-93表达；活细胞计数试剂盒（CCK-8）和流式细胞术检测miR-93对宫颈癌Hela细胞增殖及凋亡的影响；Transwell小室法检测其对Hela细胞迁移及侵袭能力的影响；Western blot检测Hela细胞中上皮生长因子受体（EGFR）及其下游蛋白的表达量变化。结果  41例宫颈癌患者肿瘤组织中miR-93表达量（0.048±0.013）低于癌旁组织（0.113±0.025），差异有统计学意义（P =0.026）；转染miR-93模拟片段后，宫颈癌Hela细胞中miR-93表达恢复；与对照组和空白组比较，实验组宫颈癌细胞增殖能力下降（P =0.004），早期凋亡比例增加（P =0.032），侵袭（P =0.003）和迁移能力下降（P =0.003）；过表达miR-93的Hela细胞EGFR及下游的p-AKT表达下降（P =0.005），而总AKT蛋白无明显变化（P =0.372）。结论  miR-93在宫颈癌组织中的表达量下降，恢复其在宫颈癌细胞中的表达能够明显抑制其细胞增殖、迁移和侵袭，促进细胞凋亡，其机制部分与miR-93抑制宫颈癌细胞EGFR/AKT信号通路活性有关。

Abstract:

Objective To investigate the expression level of miR-93 in cervical cancer tissues and its effects on the biological functions of the cancer cells. Methods Forty-one paired cervical cancer tissues and adjacent normal tissues were collected from Xiangya Hospital of Central South University from Jan. 2014 to Jul. 2014, and all the collected tissues had complete medical records. Using qRT-PCR, the expression of miR-93 was explored in the 41 paired cervical cancer tissues and adjacent normal tissues. miR-93 mimic was transfected into cervical cancer Hela cells to restore the expression of miR-93. The proliferation and apoptosis effects of miR-93 on Hela cells were evaluated by cell counting kit-8 (CCK-8) and flow cytometry, respectively. Moreover, the migration and invasion of transfected Hela cells were detected by Transwell chamber assay. Western blot was used to measure the expressions of EGFR and its downstream protein in the Hela cells. Results The expression of miR-93 was significantly decreased in the 41 cervical cancer tissues compared with the adjacent normal tissues [(0.048 ± 0.013) vs (0.113 ± 0.025), P =0.026]. By transfection of miR-93 mimic fragment, miR-93 expression recovered in the cervical cancer Hela cells. Overexpression of miR-93 through exogenous transfection with miR-93 mimic significantly suppressed cell proliferation (P = 0.004), induced cell apoptosis (P = 0.032), and inhibited cell migration and invasion (P = 0.003). Moreover, miR-93 overexpression of Hela cells significantly suppressed the expressions of EGFR and its downstream p-AKT (P = 0.005), while did not influence the expression of total AKT (P = 0.372), which suggested that overexpression of miR-93 significantly suppressed the EGFR/AKT signaling pathway in cervical cancer cells. Conclusions The expression of miR-93 is significantly decreased in cervical cancer tissues. Restored expression of miR-93 could suppress cell proliferation, migration and invasion, and induce cell apoptosis through, at least partially, suppression of EGFR/AKT signaling pathway.