慢病毒介导的P27RF-Rho基因敲低抑制肝癌细胞生长。
Lentivirus-mediated knockdown of P27RF-Rho inhibits hepatocellular carcinoma cell growth.
作者信息
Xie Shuli, Wang Guangyi, Chen Guofu, Zhu Mingguang, Lv Guoyue
机构信息
Department of General Surgery, The First Hospital of Jilin University, Jilin, China.
Department of General Surgery, Daqing City People's Hospital, Daqing City, Heilongjiang Province, China.
出版信息
Contemp Oncol (Pozn). 2017;21(1):35-41. doi: 10.5114/wo.2017.66654. Epub 2017 Mar 22.
AIM OF THE STUDY
To investigate the effects of P27RF-Rho on hepatocellular carcinoma (HCC) cell growth and explore the possibility of using it as a novel therapeutic target for liver cancer treatment.
MATERIAL AND METHODS
P27RF-Rho in HCC cells was silenced by lentivirus-mediated RNA interference, and the silencing effect was verified by RT-PCR. Cell proliferation was determined by MTT and clone formation assay. Cell cycle phase and apoptosis were detected through FACS. The expression level of cell growth, apoptosis, and metastasis associated genes was detected by quantitative PCR.
RESULTS
Lentivirus-mediated P27RF-Rho knockdown inhibited HCC cell growth and clone formation. P27RF-Rho silence induced cell cycle arrest and apoptosis. The mRNA level of genes associated with cell cycle, apoptosis, and invasion also significantly altered after P27RF-Rho knockdown. Cyclin A, CDK2, BCL-2, and MMP-9 were down-regulated. P27 and Bax were up-regulated.
CONCLUSIONS
P27RF-Rho knockdown inhibits HCC cell growth, and P27RF-Rho is probably a promising target for HCC treatment.
研究目的
探讨P27RF-Rho对肝癌（HCC）细胞生长的影响，并探索将其作为肝癌治疗新靶点的可能性。
材料与方法
采用慢病毒介导的RNA干扰技术沉默肝癌细胞中的P27RF-Rho，通过RT-PCR验证沉默效果。采用MTT法和克隆形成试验检测细胞增殖。通过流式细胞术检测细胞周期阶段和凋亡情况。采用定量PCR检测细胞生长、凋亡和转移相关基因的表达水平。
结果
慢病毒介导的P27RF-Rho基因敲低抑制了肝癌细胞的生长和克隆形成。P27RF-Rho沉默诱导细胞周期阻滞和凋亡。P27RF-Rho基因敲低后，细胞周期、凋亡和侵袭相关基因的mRNA水平也发生了显著变化。细胞周期蛋白A、细胞周期蛋白依赖性激酶2、B细胞淋巴瘤-2和基质金属蛋白酶-9表达下调。P27和Bax表达上调。
结论
P27RF-Rho基因敲低抑制肝癌细胞生长，P27RF-Rho可能是肝癌治疗的一个有前景的靶点。
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