miR-638用于治疗乙肝相关肝癌可能性的实验研究

Experimental study on the possibility of miR-638 for treatment of hepatitis B-related hepatocellular carcinoma

目的:探讨miR-638用于治疗乙肝相关肝癌的可能性。方法:qRT-PCR方法检测miR-638在乙肝、肝癌患者及正常人群血清中的表达差异,并分析血清miR-638的表达水平与其它血清指标的相关性。上调乙肝病毒(HBV)稳定复制的Hep G2.2.15肝癌细胞株中miR-638的表达水平,MTT法和细胞克隆实验检测miR-638过表达对Hep G2.2.15细胞增殖的影响;ELISA和qRT-PCR方法检测细胞培养上清液中HBsAg、HBeAg和HBV DNA的表达水平。结果:miR-638在乙肝及肝癌患者血清中的表达水平均显著低于正常人群(P<0.000 1);在乙肝及肝癌患者血清中miR-638的表达水平,总体间与HBV DNA、谷草转氨酶(AST)及谷丙转氨酶(ALT)水平呈显著负相关(P值分别为0.026、<0.001和0.003)。MTT分析及集落形成实验显示miR-638能够显著抑制Hep G2.2.15细胞的增殖及集落形成能力。miR-638表达上调后,HBs Ag(P<0.05)、HBeAg(P<0.05)和HBV DNA水平(P<0.001)较miR-ctrl组均显著下降。结论:miR-638在HBV复制过程中发挥作用,miR-638过表达可同时抑制Hep G2.2.15细胞的增殖能力和HBV的复制。

Objective： To explore the possibility of miR-638 for treatment of hepatitis B-related hepatocellular carcinoma（ HCC）. Methods： The expression of miR-638 in serum of patients with hepatitis B（ HBV）,HCC and normal population was detected by qRT-PCR. The correlation between serum miR-638 expression and other serum markers was analyzed. Pre-miR-638 expression plasmids were transfected into HepG2. 2. 15 cells. MTT assay and colony formation assay were used to analyze the role of miR-638 in HepG2. 2. 15 cells. ELISA and qRT-PCR were used to detect the expression of HBsAg,HBeAg and HBV DNA in the cell culture supernatant. Results： The significantly lower miR-638 expression was in serum/plasma of patients with HBV and HCC compared with health control（ P〈0. 000 1）. Lower serum/plasma miR-638 expression has negative relationship with HBV DNA,AST and ALT（ P values were 0. 026, 0. 001 and 0. 003,respectively）. MTT assay and colony formation assay showed that miR-638 could significantly inhibit the proliferation and colony-forming ability of HepG2. 2. 15 cells. After miR-638 was upregulated,HBs Ag（ P〈0. 05）,HBeAg（ P〈0. 05） and HBV DNA level（ P〈0. 001） decreased significantly compared with the control group. Conclusion： miR-638 plays a role in the process of HBV replication. Overexpression of miR-638 can inhibit the proliferation of HepG2. 2. 15 cells and HBV replication simultaneously.