王勇强　吴林波　陈帆

[摘要] 目的 探讨miR-10b在人肝癌组织中的表达及其对肝癌细胞增殖、侵袭和迁移的影响。 方法 搜集兰州军区乌鲁木齐总医院46例肝癌组织作为观察组，同期选取18例正常肝组织作为对照组，利用实时荧光定量聚合酶链式反应（qRT-PCR）方法检测受试者miR-10b的表达水平，分析miR-10b表达与肝癌组织病理特征之间的关系；利用脂质体2000向肝癌细胞系HepG2转染人工化学合成miR-10b模拟物（miR-10b mimics）、miR-10b抑制剂（miR-10b inhibitor），同时设置阴性对照组（NC+脂质体2000），采用MTT实验检测HepG2肝癌细胞增殖能力，采用Transwell实验及细胞划痕实验检测HepG2肝癌细胞侵袭、迁移能力。 结果 肝癌组miR-10b水平较对照组明显上调表达（P < 0.05），且肝癌转移组miR-10b水平明显高于未转移组（P < 0.05）；miR-10b高表达组与低表达组患者性别、年龄、肿瘤分化程度、有无肝硬化以及肿瘤数量之间比较差异无统计学意义（P > 0.05），肝癌患者miR-10b表达与肝癌是否转移以及肝癌的美国癌症联合委员会（AJCC）分期有关，且肝癌转移患者miR-10b表达高于未转移患者（P < 0.05），Ⅲ～Ⅳ期患者miR-10b表达高于Ⅰ～Ⅱ期（P < 0.05）；miR-10b过表达组HepG2细胞增殖能力明显高于miR-10b抑制组，过表达阴性对照组与抑制表达阴性对照组HepG2细胞生长趋势接近一致；miR-10b过表达组HepG2细胞侵袭、迁移能力较过表达阴性对照组均明显增强，miR-10b抑制组HepG2细胞侵袭、迁移能力较抑制表达阴性对照组明显减弱，过表达阴性对照组与抑制表达阴性对照组HepG2细胞侵袭、迁移能力基本一致。 结论 miR-10b在人肝癌组织中的表達水平明显高于正常肝组织，抑制miR-10b表达可有效降低肝癌细胞的增殖、侵袭及转移能力。

[关键词] miR-10b；肝癌；增殖；侵袭；转移

[中图分类号] R735.705 [文献标识码] A [文章编号] 1673-7210（2016）12（a）-0025-06

Effects of miR-10b on the proliferation， invasion and migration of hepatocellular carcinoma cells

WANG Yongqiang WU Linbo CHEN Fan▲

Department of Oncology， Urumqi General Hospital， Lanzhou Military Area Command， Xinjiang Uygur Autonomous Region， Urumqi 830099， China

[Abstract] Objective To investigate the expression of miR-10b in human hepatocellular carcinoma and its effect on the invasion and metastasis of hepatocellular carcinoma cells. Methods Forty-six cases of hepatocellular carcinoma in Urumqi General Hospital， Lanzhou Military Area Command were collected as the observation group， and 18 cases of normal liver tissues were selected as the control group. The expression levels of miR-10b of the subjects were detected by quantitative Real-time PCR （qRT-PCR） method， the relationship between the expression of miR-10b and pathological features of tissue of hepatocellular carcinoma was analyzed； the artificial chemical synthesis miR - 10b mimics， miR-10b inhibitor were transfected into hepatocellular cell line HepG2 by lipofectamine 2000， and the negative control group （NC + lipofectamine 2000） was set. MTT assay was used to detect the proliferation capacity of HepG2 cells， and Transwell assay and cell scratch test were used to detect the invasion， migration capacity of HepG2 cells. Results The level of miR-10b in liver cancer group was significantly higher than that of control group （P < 0.05）， and the level of miR-10b in metastatic hepatocelluar carcinoma group was significantly higher than that of non-metastasis group （P < 0.05）； the differences of gender， age， tumor differentiation， liver cirrhosis or not， and cancer number between high expression of miR-10b group and low expression of miR-10b group were not statistically significant （P > 0.05）； the expression of miR-10b in patients with liver cancer was related to the transfer or not of liver cancer and the American Joint Committee on Cancer （AJCC） stage of liver cancer， and the expression of miR-10b in patients with metastatic hepatocelluar carcinoma was higher than that of non-metastasis patients （P < 0.05）， the expression of miR-10b in patients with Ⅲ-Ⅳ stage was higher than that of patients with Ⅰ-Ⅱ stage （P < 0.05）； the proliferation capability in excessive expression of miR-10b group was higher than that in miR-10b inhibition group， the growth tendency of HepG2 cells of excessive expression negative control group and suppression expression negative control group was almost the same； the invasion， migration capacity of HepG2 cells in excessive expression of miR-10b group was stronger than that of excessive expression negative control group， the invasion， migration capacity of HepG2 cells in miR-10b inhibition group was weaker than that of suppression expression negative control group， the invasion， migration capacity of excessive expression negative control group and suppression expression negative control group was almost the same. Conclusion The expression level of miR-10b in tissue of human hepatocellular carcinoma is significantly higher than that of normal hepatic tissue. The inhibition expression of miR-10b can effectively reduce the proliferation， invasion and migration capacity of hepatocellular carcinoma cells.

[Key words] miR-10b； Hepatocellular carcinoma； Proliferation； Invasion； Migration

原发性肝癌（HCC）是全球范围内最为常见的恶性肿瘤之一[1]，其发病率及病死率在世界范围内均呈现明显上升趋势[2]。HCC早期发病无明显症状，大多数患者确诊时已为癌症进展期，其发病机制复杂，目前临床上仍缺乏有效的治疗性药物。肝癌细胞的侵袭转移是导致肝癌患者死亡的主要原因，探究肝癌细胞的侵袭转移机制对于临床治疗肝癌具有重要意义。近年来，微小RNA研究为肿瘤靶向研究提供了新的研究方向，成为近年来肿瘤研究的新热点。相关研究表明[3-4]，微小RNA成员miR-10b在多种肿瘤中呈现高表达，且对肿瘤细胞的侵袭及转移具有促进作用。……