UNASSIGNED:根据生物信息学分析,已显示WNK赖氨酸缺陷蛋白激酶1(WNK1)在肝细胞癌(HCC)样品中高表达,并与HCC患者的不良预后有关。然而,WNK1在HCC中的具体功能尚未分析。本研究旨在探讨WNK1在HCC进展中的作用及其相关分子机制。
未经鉴定:通过小干扰RNA敲低WNK1后,细胞计数试剂盒-8,集落形成,westernblot,Transwell,和伤口愈合试验用于评估HCC细胞的生物学行为。应用免疫荧光染色检测WNK1对LC3Ⅱ的影响。应用GSK690693或si-AMPK阻断AMPK途径。Westernblot检测自噬和AMPK通路相关分子的表达。
UNASSIGNED:WNK1在HCC细胞系中高表达,WNK1的缺失抑制了HCC细胞的增殖,细胞周期,迁移,和入侵。此外,我们证明WNK1的缺失促进了HCC细胞的自噬和激活的AMPK通路。同时,GSK690693处理或si-AMPK转染抑制自噬并促进HCC细胞增殖。然而,WNK1敲低抵消了GSK690693或si-AMPK在调节HCC细胞增殖中的作用。最后,我们证明WNK1通过调节自噬和AMPK通路调节肝癌细胞的恶性行为。
UNASSIGNED:上述结果表明WNK1可能是肝癌治疗值得考虑的靶点。
UNASSIGNED: WNK lysine deficient protein kinase 1 (WNK1) has been shown to be highly expressed in hepatocellular carcinoma (HCC) samples and related to poor prognosis of HCC patients based on bioinformatics analysis. However, the specific function of WNK1 in HCC has not been analyzed. This study is aimed at exploring the function of WNK1 in HCC progression as well as its related molecular mechanism.
UNASSIGNED: After knockdown of WNK1 by small interference RNA, cell counting kit-8, colony formation, western blot, Transwell, and wound healing assays were employed to evaluate the biological behaviors of HCC cells. Immunofluorescent staining was applied to detect the effect of WNK1 on LC3 II. GSK690693 or si-AMPK was applied to block AMPK pathway. The expression of autophagy and AMPK pathway related molecules was examined by western blot assay.
UNASSIGNED: WNK1 was highly expressed in HCC cell lines and loss of WNK1 inhibited HCC cell proliferation, cell cycle, migration, and invasion. Additionally, we demonstrated that loss of WNK1 promoted the autophagy and activated AMPK pathway in HCC cells. While, GSK690693 treatment or si-AMPK transfection suppressed the autophagy and promoted HCC cells proliferation. However, WNK1 knockdown counteracted the effect of GSK690693 or si-AMPK in regulating HCC cell proliferation. Finally, we demonstrated that WNK1 regulated the malignant behaviors of HCC cells by modulating autophagy and AMPK pathway.
UNASSIGNED: The above results indicated that WNK1 may be a worthwhile target to be considered for therapy of HCC.