DCK基因敲减对宫颈癌HeLa细胞增殖、侵袭及放化疗敏感性的影响  

Impact of DCK Knockdown on Proliferation, Invasion and Response to Chemoradiotherapy in Cervical Cancer HeLa Cells

作　　者：王健 吴丛山 高华荣[2] 成昌宏 尚庆毅[1] WANG Jian;WU Congshan;GAO Huarong;CHENG Changhong;SHANG Qingyi(Department of Clinical Laboratory,Renmin Hospital,Lianyungang City,Lianyungang,Jiangsu,222100,China;Department of Gynecology,Lianyungang People's Hospital,Lianyungang City,Lianyungang,Jiangsu,222100,China)

机构地区：[1]江苏连云港市赣榆区人民医院检验科,江苏省连云港市222100 [2]江苏省连云港市赣榆区人民医院妇科,江苏省连云港市222100

出　　处：《医学分子生物学杂志》2019年第6期531-536,共6页Journal of Medical Molecular Biology

基　　金：江苏省连云港市卫生和计划生育委员会(No.201836)。

摘　　要：目的 研究DCK对宫颈癌HeLa细胞增殖及侵袭活性影响, 对HeLa细胞顺铂及放疗的敏感性影响.方法 siRNA干扰DCK基因在HeLa细胞中的表达. CCK-8实验、克隆形成试验及细胞侵袭实验分别检测DCK基因敲减后细胞增殖、克隆形成及侵袭能力变化.不同浓度顺铂或不同剂量放射线处理HeLa对照及DCK敲减细胞, CCK-8检测细胞增殖活性变化.利用细胞自噬抑制剂3-甲基腺嘌呤、自噬诱导剂雷帕霉素及不同放射剂量处理HeLa对照及DCK敲减细胞, 检测细胞增殖活性及自噬活化标记 LC-Ⅱ, 揭示DCK在放射诱导细胞自噬中的作用.结果 DCK基因敲减后, HeLa细胞增殖活性及克隆形成能力显著减弱 (P<0.01);细胞侵袭能力与对照组比较也明显减弱 (P<0.01).对于顺铂处理组, 我们发现随着顺铂药物浓度增加HeLa细胞的细胞活力明显降低, 然而DCK未敲减对照组与敲减组之间的细胞活力没有显著变化 (P>0.05).对于放疗处理组, 未敲减对照组细胞随着放射剂量增加, 细胞活力下降;而DCK敲减组细胞活力与对照组比较进一步下降 (P<0.01);对于放射线处理合并DCK敲减处理组, 3-甲基腺嘌呤可导致进步抑制细胞增殖 (P<0.01), 而雷帕霉素可部分逆转DCK敲减导致的细胞增殖抑制 (P<0.01).对HeLa细胞进行低剂量 (2 Gy) 及高剂量 (6 Gy) 放射处理, 对照组细胞LC-Ⅰ/LC-Ⅱ比值相对表达显著下调 (P<0.05), 而DCK敲减组, 无论低剂量 (2 Gy) 及高剂量 (6 Gy) 放射处理, LC-I/LC-II比值都处于较低水平.结论 DCK对宫颈癌细胞增殖及转移起到重要促进作用, 同时敲减DCK后, HeLa细胞对放射敏感性增高.Objective To investigate the effect of DCK knockdown on cell proliferation, inva-sion and response to chemoradiotheray in cervical cancer HeLa cells.Methods DCK expression was reduced by siRNA in HeLa cells.CCK-8 assay, colony formation assay and invasion assay were used to evaluate the proliferation rate, colony formation and invasion ability of HeLa cells with or without treatment of DCK siRNA.Different dosage of cisplatin and X-rays were used for the treatment of control and DCK knowckdown cells, and CCK-8 assay was then used to evaluate their prolifera-tion ability.By using autophagy inhibitor 3-methyladenine and inducer rapamycin, and different dos-age of X-rays, CCK-8 assay and westernblot assay were used to measure proliferation rate and LC-Ⅱ expression in control and DCK knockdown cells.Results The proliferation rate, colony forma- tion and invasion ability was significant inhibited by DCK knockdown ( P<0.01);For the cispla-tin treatment, the proliferation of control cells significant decreased in a dose dependent manner, however there is no different of growth rate between control and DCK knockdown cells ( P>0.05) . For the X-ray treatment, the proliferation rate of control cells significant decreased in a dose de-pendent manner, and the growth rate is further inhibited in DCK knockdown cells compared to con-trol cells (P<0.01) .For the treatment of X-rays and DCK knockdown, 3-methyladenine caused further growth inhibitory ( P<0.01) , while rapamycin is able to reverse growth inhibitory induced by DCK knockdown ( P<0.01) .For the treatment of 2 Gy and 6 Gy of X-rays, the expression level of LC-Ⅱ is significantly induced in control cells, while no obvious change was observed in DCK knockdown cells.Conclusion DCK plays an important role in proliferation and invasion in cervical cancer cells.Knockdown of DCK caused increased response to radiotherapy in cervical canc-er.

关 键 词：宫颈癌 DCK 顺铂 放射 细胞自噬 

分 类 号：R711.7[医药卫生—妇产科学]