RNA干扰抑制JMJD3基因表达对肝癌细胞生长,增殖和侵袭的影响

Influence of RNAi inhibiting JMJD3 gene expression on growth,proliferation and invasion of hepatocellular carcinoma cells

目的研究小干扰RNA(siRNA)抑制含Jumonji结构域蛋白3(JMJD3)基因对肝癌细胞QGY-7703和HepG2的生长、增殖和侵袭能力的影响。方法利用RNA干扰技术沉默JMJD3(siR-JMJD3),以非特异性序列(pSilencer 2.1)转染肝癌细胞QGY-7703细胞和HepG2细胞作为阴性对照,采用四甲基噻唑蓝(MTT)法、集落形成、Transwell侵袭实验来检测肝癌细胞生长、增殖和侵袭能力的变化。结果 Western Blot检测结果显示,转染siR-JMJD3质粒的试验组成功抑制肝癌细胞中JMJD3的蛋白表达水平。MTT结果显示转入siR-JMJD3后,QGY-7703和HepG2细胞的生长活性与对照组相比分别降低了约25%和17%。集落形成结果显示2种细胞系的集落形成数分别降低了约31%和25%。Transwell侵袭实验结果显示穿膜细胞数分别下降了约44%和47%。结论应用siRNA技术能有效抑制JMJD3基因的表达,同时有效抑制肝癌细胞QGY-7703和HepG2体外生长、增殖和侵袭,为肝癌的生物学治疗提供了新思路。

Objective To study the influence of small interfering RN A （siRNA） inhibiting Jumonji-containing JMJD3 gene on growth, proliferation and invasion of hepatocellular carcinoma cells. Methods JMJD3 was silenced by RNA interference （siR- JMJD3） , non-specificity sequence pSilercer 2. 1 was transfected into liver cancer cells QGY 7703 and HepG2 cells as the negative control. MTT assay,colony formation and Transwell invasion assays were used to detect the cell growth,proliferation and invasion ability of hepatocellular carcinoma cells. Results The Western Blot detection results showed that the siR- JMJD3 plasmid transfect-ed experiment group successfully inhibited the JMJD3 protein expression level in liver cancer cells. The MTT results showed that after transfecting siR- JMJD3 ,the growth activity of QGY-7703 and HepG2 were decreased by 25% or 17% compared with the control group,. The colony formation results showed that in the colony formation number of two cell lines were decreased by 31% and 25% respectively. The Transwell invasion test results indicated that transmembrane cells number was decreased by 44 % and 47% respectively. Conclusion Using siRNA technique can effectively inhibit JMJD3 gene expression, meanwhile effectively inhibits the in vitro growth, proliferation and invasion of liver cancer cells QGY-7703 and HepG2 and provides a new thinking for the biological therapy of liver cancer.