| 摘要: |
| 目的观察RECQL基因沉默表达对人肝细胞癌(humanhepatocelullarcarcinoma,HCC)细胞株体外增殖的影响,探讨RECQL作为肝癌治疗潜在靶向基因的可能性。方法以BEL7404、SK-HEP-1、Hep3B、SNU-761HCC细胞为研究对象，采用Westernblot和免疫组化法观察解旋酶RECQL蛋白的表达情况，通过RNA干扰技术抑制RECQL表达，测定HCC细胞和正常肝细胞（THLE-2细胞）的存活率和死亡率，并采用Westernblot法分析凋亡相关蛋白的变化，FACS法检测细胞周期分布。结果RECQL蛋白在BEL7404、SK-HEP-1、Hep3B中为阳性表达，SNU-761中为阴性表达。RECQL-siRNA在阳性表达组中抑制HCC细胞的增殖，引起细胞存活率降低和死亡率增加（均P＜0.01），但对阴性表达组和THLE-2细胞存活率和死亡率无影响（均P＞0.05）。RECQL-siRNA处理的SK-HEP-1细胞中，剪切型凋亡相关蛋白cleavedcaspase-3和cleavedPARP的表达变化不大，而G2/M期细胞的比例明显增加（P＜0.05或0.01）。结论RECQL基因沉默表达可以抑制HCC细胞的增殖，且这种抑制作用依赖于RECQL的高表达，并对正常肝细胞无影响，其可能机制是阻滞细胞G2/M期，可以作为治疗肝癌的潜在靶向基因。 |
| 关键词: RECQL 人肝细胞癌 细胞凋亡 细胞周期 |
| DOI：10.12056/j.issn.1006-2785.2017.39.05.2016-1804 |
| 分类号: |
| 基金项目:国家自然科学基金项目（81602671）；浙江省自然科学基金项目（LQ17H160003） |
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| RECQL-siRNA inhibits proliferation of human hepatocellular carcinoma cell lines in vitro |
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ZHANG Jingjing, ZHAO Yanyan, XU Yasi, HUANG Haixiu, ZHU Lucheng, MA Shenglin, ZHANG Shirong
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Hangzhou Hospital of Nanjing Medical University
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| Abstract: |
| Objective To investigate the effect of RECQL on proliferation of human hepatocellular carcinoma cell lines in vitro. Methods Hepatocellular carcinoma (HCC) BEL7404, SK-HEP-1, Hep3B and SNU-761 cells were transfected with RECQL-siRNA in vitro to silence RECQL. The expression of RECQL protein was detected with Western blot and immunohistochemistry. The viability of HCC and normal liver THLE-2 cells were examined, the expressions of apoptosis-related proteins were detected with Western blot and cell cycle distribution was observed using FACS. Results RECQL protein was positive in BEL7404, SK-HEP-1 and Hep3B cells, and negative in SNU-761 cells. RECQL-siRNA inhibited the proliferation of
HCC cells significantly in the RECQL-positive expression HCC cells (P＜0.05), but had no effect on HCC SNU-761 cells and
normal liver THLE-2 cells. In SK-HEP-1 cells transfected with RECQL-siRNA, the expression of cleaved caspase-3 and cleaved PARP had no significant change, but the ratio of G2/M phase cells was significantly increased (P ＜0.05 or 0.01). Conclusion RECQL-siRNA can inhibit the proliferation of HCC cells with positive RECQL, indicating that RECQL may be a potential target gene for treatment of liver cancer. |
| Key words: RECQL Human hepatocellular carcinoma Cell apoptosis Cell cycle |
