KIF14在肝细胞癌SMCC-7721获得性耐药机制中的研究

目的 探究抑制KIF14与肝细胞癌SMCC-7721对索拉非尼获得性耐药的关系。方法 采用浓度梯度法建立肝细胞癌索拉非尼耐药细胞系SMCC-7721-SR,后采用CCK-8法、流式细胞仪验证耐药细胞株的耐药性。在人正常肝细胞LO2、肝细胞癌SMCC-7721及其耐药细胞SMCC-7721-SR中,采用qRT-PCR及Western blot法检测KIF14的表达情况。随后使用KIF14特异性的小干扰RNA(siKIF14)敲低KIF14并通过Western blot法检测KIF14的表达;为探究敲低KIF14后联合索拉非尼对肝细胞癌索拉非尼耐药细胞的影响,采用CCK-8法检测细胞活力、流式细胞仪检测细胞凋亡。结果 与LO2相比SMCC-7721中KIF14表达增高(P＜0.01),与SMCC-7721相比SMCC-7721-SR中KIF14表达也增强(P＜0.01);敲低KIF14的表达可协同索拉非尼抑制SMCC-7721-SR的细胞活力、促进细胞凋亡(P＜0.01)。结论 KIF14参与调控SMCC-7721索拉非尼的获得性耐药,抑制KIF14的表达可增强肝细胞癌SMCC-7721-SR对索拉非尼的敏感性。

KIF14 in the mechanism of acquired drug resistance in hepatocellular carcinoma SMCC-7721 cells

Objective The objective of this study was to explore the relationship between inhibition of KIF14 and acquired resistance of hepatocellular carcinoma(HCC)SMCC-7721 cells to sorafenib.Methods The concentration gradient method was used to establish the HCC sorafenib-resistant SMCC-7721-SR cell line,and then the CCK-8 method and flow cytometry were used to verify resistant cell properties of the drug-resistant cell line.The qRT-PCR and Western blot assays were used to determine the expression of KIF14 in human normal liver LO2 cells,HCC SMCC-7721 cells and drug-resistant SMCC-7721-SR cells.Subsequently,KIF14-specific small interfering RNA(siKIF14)was used to knock down KIF14 and the KIF14 level was verified by Western blot;in order to explore the effect of knocking down KIF14 combined with sorafenib on HCC sorafenib-resistant cells,the CCK-8 method was used to detect cell viability,and flow cytometry was used to detect cell apoptosis.Results Compared with LO2 cells,the expression of KIF14 in SMCC-7721 cells was increased(P<0.01),and the expression of KIF14 in SMCC-7721-SR cells was also increased when compared with SMCC-7721 cells(P<0.01).Knocking down KIF14 expression could synergistically sorafenib inhibit the cell viability of SMCC-7721-SR cells and promote cell apoptosis(P<0.01).Conclusion KIF14 is involved in the process of HCC and regulates the acquired sorafenib resistance of SMCC-7721 cells.Inhibiting the KIF14 expression could enhance the sensitivity of SMCC-7721-SR cells to sorafenib.