Liu Jiangwei, Li Xiaocheng, Huang Jianzhao, Liu Yan
Department of Hepatobiliary Surgery, Guizhou Provincial People's Hospital, No. 1 Baoshan South Road, Guizhou 550002, China.
Open Med (Wars). 2019 May 17;14:384-391. doi: 10.1515/med-2019-0039. eCollection 2019.
This study evaluated the functions of matrix metalloproteinase 2 (MMP2) in hepatocellular carcinoma (HCC) cells and assessed the effects of MMP2 on HCC cell sensitivity to cisplatin.
HepG2 and Huh7 cells were cultured. A pre-experiment was performed to explore the optimal transduction conditions of the MMP2-siRNA lentivirus (si-MMP2). Quantitative real-time PCR and western blot assays were performed to measure the expression levels of MMP2 in HepG2 and Huh7 cells. An MTT assay was used to evaluate cell proliferation, and flow cytometry analysis was applied to examine cell apoptosis. A Transwell assay was carried out to assess cell invasion.
The optimal virus:cell ratio was 100 multiplicity of infection (MOI) for both cells, and the optimal transduction times for HepG2 and Huh7 cells were 48 h and 72 h, respectively. MMP2 knockdown significantly decreased the mRNA and protein levels of MMP2 in both cell lines (P<0.01). MMP2 knockdown significantly decreased the proliferation and increased the apoptosis of HepG2 and Huh7 cells (P<0.01). Co-treatment with si-MMP2 and cisplatin significantly increased the sensitivity of HepG2 and Huh7 cells to cisplatin (P<0.01).
MMP2 may act as an oncogene and may be a potential therapeutic target in HCC.
本研究评估了基质金属蛋白酶2(MMP2)在肝癌(HCC)细胞中的功能,并评估了MMP2对HCC细胞顺铂敏感性的影响。
培养HepG2和Huh7细胞。进行预实验以探索MMP2-siRNA慢病毒(si-MMP2)的最佳转导条件。采用定量实时PCR和蛋白质印迹分析来检测HepG2和Huh7细胞中MMP2的表达水平。使用MTT法评估细胞增殖,并应用流式细胞术分析检测细胞凋亡。进行Transwell实验以评估细胞侵袭。
两种细胞的最佳病毒:细胞比例均为100感染复数(MOI),HepG2和Huh7细胞的最佳转导时间分别为48小时和72小时。MMP2基因敲低显著降低了两种细胞系中MMP2的mRNA和蛋白质水平(P<0.01)。MMP2基因敲低显著降低了HepG2和Huh7细胞的增殖并增加了其凋亡(P<0.01)。si-MMP2与顺铂联合处理显著增加了HepG2和Huh7细胞对顺铂的敏感性(P<0.01)。
MMP2可能作为一种癌基因,可能是HCC潜在的治疗靶点。