摘要
[目的]研究CHD1L调节PI3K通路对皮肤鳞状细胞癌(Cutaneous Squamous Cell Carcinoma,CSCC)A431细胞增殖、凋亡的影响。[方法]将A431细胞吹打均匀。将A431细胞分为3组(2×10^(6)个细胞/组):阴性对照组(转染0.1μmol/L siRNA NC)、siCHD1L组(转染0.1μmol/L siRNA CHD1L)、LY294002组(加入5μmol/L PI3K通路抑制剂)。以蛋白免疫印迹实验分析CSCC细胞(A431、SCC13、HSC-5)、正常皮肤细胞(HaCaT)中CHD1L的表达。采用MTT实验、细胞集落形成实验分析A431细胞增殖活性;原位末端标记法分析A431细胞凋亡率;蛋白免疫印迹实验检测A431细胞中CHD1L、PI3K与AKT蛋白的表达水平。[结果]与正常皮肤细胞相比,CSCC细胞中的CHD1L表达水平增加(P<0.05)。与阴性对照组相比,siCHD1L组、LY294002组的A431细胞的增殖能力降低(P<0.05)、凋亡率增加(P<0.05)。与阴性对照组相比,siCHD1L组的CHD1L、PI3K和AKT表达降低(P<0.05),LY294002组的PI3K与AKT表达降低,而LY294002组的CHD1L表达水平无明显变化。[结论]CSCC细胞中CHD1L表达量高于正常皮肤细胞,并且抑制CSCC A431细胞中的CHD1L后,PI3K通路同时受到抑制。通过抑制CHD1L或PI3K通路能够降低CSCC A431细胞的增殖活性,并促其凋亡。
[Objective]To investigate the effect of CHD1L on the proliferation and apoptosis of cutaneous squamous cell carci⁃noma(CSCC)A431 cells by regulating PI3K pathway.[Method]Divide the A431 cells into 3 groups(2×10^(6)A431 cells per group):negative control group(transfected with 0.1μmol/L siRNA NC),siCHD1L group(transfected with 0.1μmol/L siRNA CHD1L),and LY294002 group(treated with 5μmol/L PI3K pathway inhibitor).The expression of CHD1L in skin squamous cell carcinoma cells(A431,SCC13,HSC-5)and normal skin cells(HaCaT)was analyzed by Western blot.The proliferation activity of A431 cells was analyzed by MTT assay and colony formation assay;the apoptosis rate of A431 cells was analyzed by in situ terminal labeling;and the expression levels of CHD1L,PI3K and AKT proteins in A431 cells were detected by Western blot.[Result]CHD1L expression levels were increased in CSCC compared with normal skin cells(P<0.05).Compared with the negative control group,the proliferation ability of A431 cells in the siCHD1L group and the LY294002 group was decreased(P<0.05),and the apoptosis rate was increased(P<0.05).Compared with the negative control group,the expression of CHD1L,PI3K and AKT in the siCHD1L group was decreased(P<0.05),while the expression of PI3K and AKT in the LY294002 group was decreased,and the expression level of CHD1L in the LY294002 group showed no significant change.[Conclusion]Compared with normal skin cells,the expression level of CHD1L was increased in CSCC,and inhibition of CHD1L in CSCC A431 cells inhibited the PI3K pathway.Inhibition of CHD1L or PI3K pathway can reduce the proliferation activity of A431 cells and promote their apoptosis.
作者
董海平
吕超
李梓琰
刘梅
DONG Haiping;LYU Chao;LI Ziyan;LIU Mei(Department of Oncology,Handan First Hospital,Handan 056000;Department of Dermatology,Affiliated Hospital of Hebei Engineering University,Handan 056000,China)
出处
《生物技术》
2025年第6期744-748,共5页
Biotechnology
基金
河北省医学科学研究课题计划资助(20240523)。
