摘要
目的通过分子生物学技术手段实现趋化素样因子1(CKLF1)在肾癌细胞系中的过表达,并探讨CKLF1对肾癌细胞增殖、迁移和侵袭活性的影响及相关机制。方法使用购自中国科学院典型培养物保藏委员会细胞库的人ACHN细胞系作为研究模型,通过转染CKLF1质粒到细胞中来实现CKLF1的过表达。利用IncuCyte S3活细胞动态成像系统观察细胞增殖。Transwell法评估细胞迁移和侵袭能力,最后进行结晶紫染色和显微镜下计数来评估。两组间比较采用独立样本t检验。结果转染CKLF1质粒后,在0 h时对照组细胞密度(182.93±80.42)与观察组(178.23±68.23)比较,差异无统计学意义(t=0.077,P>0.05);在48 h时,对照组细胞密度(892.52±204.34)明显低于观察组(1974.23±382.42),差异有统计学意义(t=4.314,P<0.05)。Transwell法的结果显示,1 h时对照组迁移细胞百分比[(22±3)%]与观察组[(23±2)%]比较,差异无统计学意义(t=0.481,P>0.05);在8 h时,对照组迁移细胞百分比[(29±3)%]明显低于观察组[(59±5)%],差异有统计学意义(t=8.911,P<0.05)。而通过对侵袭能力进行测定显示,1 h时对照组侵袭细胞百分比[(21±3)%]与观察组[(19±2)%]比较,差异无统计学意义(t=0.961,P>0.05);在8 h时,对照组侵袭细胞百分比[(26±5)%]明显低于观察组[(55±4)%],差异有统计学意义(t=7.845,P<0.05)。结论CKLF1在肾癌细胞中具有促进增殖、迁移和侵袭能力的作用。
ObjectiveTo achieve the overexpression of chemokine-like factor 1(CKLF1)in renal cancer cell lines through molecular biology techniques,and to explore the effects and related mechanisms of CKLF1 on the proliferation,migration and invasion activities of renal cancer cells.MethodsThe ACHN cell line was used as the study model to achieve overexpression of CKLF1 by transfecting CKLF1 plasmid carrying purinamycin resistance into cells,and then the cells were screened by purinamycin.IncuCyte S3 live cell dynamic imaging system was used to observe cell proliferation in CKLF1 overexpression group and control group.Transwell method was used to evaluate the cell migration and invasion ability of CKLF1 overexpression group and control group.Finally,the cells were stained with crystal violet and counted under microscope for further comparison.Independent sample t-test was used for comparison between the two groups.ResultsOverexpression of CKLF1 significantly promoted the proliferation of renal carcinoma cells.After transfection with CKLF1,there was no significant difference in cell density between control group(182.93±80.42)and observation group(178.23±68.23)at 0 h(t=0.077,P>0.05).At 48 h,the cell density of the control group(892.52±204.34)was significantly lower than that of the observation group(1974.23±382.42,t=4.314,P<0.05).Overexpression of CKLF1 also significantly increased the migration ability of renal cancer cells.The results of Transwell method showed that there was no significant difference in the percentage of migrating cells between the control group[(22±3)%]and the observation group[(23±2)%]at 1 h(t=0.481,P>0.05).At 8 h,the percentage of migrating cells in control group[(29±3)%]was significantly lower than that in observation group[(59±5)%,t=8.911,P<0.05].The invasion ability of the control group[(21±3)%]and the observation group[(19±2)%]at 1 h showed no significant difference(t=0.961,P>0.05).At 8 h,the percentage of invasive cells in the control group[(26±5)%]was significantly lower than that in the
作者
骆杨
胡伟
杨富斌
Luo Yang;Hu Wei;Yang Fubin(The Fifth Department of Surgical Oncology,Xinyang Central Hospital,Xinyang 464000,China)
出处
《中华实验外科杂志》
CAS
2024年第8期1763-1766,共4页
Chinese Journal of Experimental Surgery
关键词
趋化素样因子1
肾癌
肿瘤侵袭
Chemokine-like factor 1
Renal cancer
Tumor invasion
