CDKN3对宫颈癌细胞增殖及侵袭迁移能力的影响  

Effect of CDKN3 on Proliferation,Invasion and Migration Ability of Cervical Cancer Cells

作　　者：郑乐[1] 张俭[1] 陆瑜珊 贾海清 ZHENG Le;ZHANG Jian;LU Yushan;JIA Haiqing(Department of Gynecology,Cancer Hospital of Dalian University of Technology,Liaoning Cancer Hospital and Institute,Shenyang Liaoning 110042,China)

机构地区：[1]大连理工大学附属肿瘤医院·辽宁省肿瘤医院妇科,辽宁沈阳110042

出　　处：《联勤军事医学》2024年第11期911-915,951,共6页Military Medicine of Joint Logistics

基　　金：国家自然科学基金资助项目(81572054)。

摘　　要：目的 探讨细胞周期蛋白依赖性激酶抑制剂3(cyclin-dependent kinase inhibitor 3,CDKN3)的表达对宫颈癌细胞增殖、侵袭和迁移能力的影响。方法 本实验分为HeLa细胞株CDKN3 siRNA组(沉默CDKN3 siRNA)和阴性对照组(未干扰CDKN3 siRNA)、SiHa细胞株CDKN3 siRNA组(沉默CDKN3 siRNA)和阴性对照组(未干扰CDKN3 siRNA)。通过免疫荧光法检测宫颈癌细胞系中的CDKN3蛋白;采用Western blot检测CDKN3在宫颈癌HeLa、SiHa细胞中的表达水平;采用细胞计数试剂盒(cell counting kit 8,CCK-8)法检测转染siRNA后的宫颈癌细胞增殖能力情况;Transwell小室检测转染siRNA后的宫颈癌细胞侵袭能力情况;细胞划痕试验检测转染siRNA后的宫颈癌细胞迁移能力情况。结果 荧光显微镜下,CDKN3蛋白表达主要定位于宫颈癌细胞的细胞核,部分表达于细胞质。Western blot结果显示,CDKN3在宫颈癌细胞中呈高表达(P<0.05)。通过CCK-8法检测发现,siRNA干扰后SiHa、HeLa细胞CDKN3 siRNA组及阴性对照组的宫颈癌细胞增殖能力从第1天开始增强(P均<0.05),且从第2天开始CDKN3 siRNA组细胞增殖能力明显小于阴性对照组(P均<0.05)。Transwell试验及细胞划痕试验中,宫颈癌HeLa和SiHa细胞中CDKN3基因沉默导致侵袭细胞数量、迁移细胞数量显著减少(P均<0.05)。结论 沉默CDKN3可抑制宫颈癌HeLa、SiHa细胞增殖、侵袭、迁移能力,未来可能会成为判断宫颈癌的预后参考指标和治疗靶点。Objective To investigate the effects of the expression of cyclin-dependent kinase inhibitor 3(CDKN3) on proliferation,invasion,and migration of cervical cancer cells.Methods The HeLa,SiHa cell line were divided into CDKN3 siRNA group(silent CDKN3 siRNA) and negative control group(no interference with CDKN3 siRNA),respectively.CDKN3 protein in cervical cancer cell line was detected by immunofluorescence assay;Western blot was used to detect the expression level of CDKN3 in HeLa and SiHa cells of cervical cancer;cell counting kit 8(CCK-8) was used to detect the proliferation ability of cervical cancer cells after transfection with siRNA;Transwell chamber was used to detect the invasion ability of cervical cancer cells transfected with siRNA;Cell scratch assay was used to detect the migration ability of cervical cancer cells transfected with siRNA.Results Under fluorescence microscopy,CDKN3 protein expression was mainly localized in the nucleus of cervical cancer cells,and some expressed in the cytoplasm;Western blot analysis showed that CDKN3 was highly expressed in cervical cancer cells(P<0.05);CCK-8 assay showed that the proliferation ability of SiHa cells and HeLa cells of cervical cancer after siRNA interference in CDKN3 siRNA group and negative control group began to increase from the 1st day(P < 0.05),and the proliferation ability of CDKN3 siRNA group was significantly lower than that of negative control group from the second day(P < 0.05).Transwell assay and cell scratch assay showed that CDKN3 gene silencing in HeLa and SiHa cells significantly reduced the number of invasive and migrating cells(P < 0.05).Conclusion Silencing CDKN3 can inhibit the proliferation,invasion and migration of HeLa and SiHa cells of cervical cancer,and may become a prognostic reference and treatment target for cervical cancer in the future.

关 键 词：细胞周期蛋白依赖性激酶抑制剂3 宫颈癌 增殖 侵袭 迁移 

分 类 号：R737.33[医药卫生—肿瘤]