核不均一核糖核蛋白L对口腔鳞状细胞癌细胞增殖、迁移和侵袭的影响  

The effect of heterogeneous nuclear ribonucleoprotein L on the proliferation,migrate,and invasion of oral squamous cell cancer cells

作　　者：刘纪雷 谢志文[1] 郭莉莉 袁晓琳 唐洁 韩小幸[1] 刘庆庆 魏士刚[1] 付东杰[2] Liu Jilei;Xie Zhiwen;Guo Lili;Yuan Xiaolin;Tang Jie;Han Xiaoxing;Liu Qingqing;Wei Shigang;Fu Dongjie(Department of Stomatology,the First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Department of Stomatology,People’s Hospital of Wuhan University,Wuhan 430000,China)

机构地区：[1]河南中医药大学第一附属医院口腔外科,郑州450000 [2]武汉大学人民医院口腔科,武汉430000

出　　处：《中华实验外科杂志》2023年第11期2292-2295,共4页Chinese Journal of Experimental Surgery

基　　金：湖北省卫生与计划委员会自然科学基金(WJ2017F001)。

摘　　要：目的:探讨核不均一核糖核蛋白L(hnRNPL)对口腔鳞状细胞癌细胞增殖、迁移和侵袭的影响。方法:采用蛋白质免疫印迹(Western blot)和荧光定量聚合酶链反应(PCR)分析正常口腔上皮细胞(HOK)和口腔鳞状细胞癌细胞系(HSC3、CAL27、SCC15、HN13)中hnRNPL蛋白和信使RNA(mRNA)表达水平;以CAL27细胞作为研究对象,采用对照短发卡RNA(shRNA)和hnRNPL shRNA慢病毒感染CAL27细胞,建立对照组和hnRNPL KD组细胞,采用细胞计数试剂盒(CCK-8)和EdU染色法检测两组细胞的增殖能力;采用流式细胞术分析两组细胞的凋亡水平;采用划痕实验和Transwell分析两组细胞的迁移和侵袭能力;采用Western blot和荧光定量PCR分析两组细胞p53、B细胞淋巴瘤/白血病-2(bcl-2)和黏着斑激酶(FAK)蛋白和mRNA表达水平。组间比较采用t检验。结果:口腔鳞状细胞癌细胞系(HSC3、CAL27、SCC15、HN13)hnRNPL蛋白表达水平(1.40±0.12、1.81±0.16、1.38±0.09、1.39±0.10)明显高于正常口腔上皮细胞(HOK,0.97±0.12),差异有统计学意义(t=6.451、10.230、6.729、6.707,P<0.05)。口腔鳞状细胞癌细胞系(HSC3、CAL27、SCC15、HN13)hnRNPL mRNA表达水平(1.47±0.13、2.04±0.22、1.59±0.10、1.42±0.08)明显高于正常口腔上皮细胞(HOK,0.99±0.05),差异有统计学意义(t=8.307、11.260、13.820、11.560,P<0.05)。对照组细胞吸光度(A)值(2.04±0.13)明显高于hnRNPL KD组细胞(1.61±0.11),差异有统计学意义(t=6.322,P<0.05)。对照组细胞EdU阳性率[(70.47±10.42)%]明显高于hnRNPL KD组[(43.71±4.54)%],差异有统计学意义(t=5.765,P<0.05)。对照组细胞凋亡率[(3.72±1.10)%]明显低于hnRNPL KD组[(11.81±2.43)%],差异有统计学意义(t=7.427,P<0.05)。对照组细胞愈合率[(86.55±4.66)%]明显高于hnRNPL KD组[(57.34±6.20)%],差异有统计学意义(t=9.224,P<0.05)。对照组细胞迁移数量[(106.50±7.66)个]明显高于hnRNPL KD组[(77.83±6.18)个],差异有统计学意义(t=7.135,P<0.05)。对照组细胞p53和bcObjective To investigate the effects of heterogeneous nuclear ribonucleoprotein L(hnRNPL)on the proliferation,migration,and invasion of oral squamous cell carcinoma cells.Methods The expression levels of hnRNPL protein and mRNA in normal oral epithelial cells(HOK)and oral squamous cell carcinoma cell lines(HSC3,CAL27,SCC15,HN13)were analyzed by Western blotting and fluorescence quantitative polymerase chain reaction(PCR).Using CAL27 cells as the research object,control short hairpin RNA(shRNA)and hnRNPL shRNA lentivirus were used to infect CAL27 cells,and control group and hnRNPL KD group cells were established.The proliferation ability of the two groups of cells was detected using cell counting kit-8(CCK-8)and EdU staining.The apoptosis levels of the two groups of cells was analyzed by flow cytometry.The migration and invasion abilities of the two groups of cells were analyzed using scratch experiments and Transwell analysis.The expression levels of p53,B cell lymphoma/leukemia-2(bcl-2),and focal adhesion kinase(FAK)proteins and mRNA in the two groups of cells were analyzed by Western blotting and fluorescence quantitative PCR.The comparison of measurement data between groups was conducted using t-test.Results The expression levels of hnRNPL protein in oral squamous cell carcinoma cell lines(HSC3,CAL27,SCC15,HN13)(1.40±0.12,1.81±0.16,1.38±0.09,1.39±0.10)were significantly higher than those in normal oral epithelial cells(HOK)(0.97±0.12.t=6.451,10.230,6.729,6.707,P<0.05).The expression level of hnRNPL mRNA in oral squamous cell carcinoma cell lines(HSC3,CAL27,SCC15,HN13)(1.47±0.13,2.04±0.22,1.59±0.10,1.42±0.08)was significantly higher than that in normal oral epithelial cells(HOK)(0.99±0.05.t=8.307,11.260,13.820,11.560,P<0.05).The absorbance value(A)of the control group cells(2.04±0.13)was significantly higher than that of the hnRNPL KD group cells(1.61±0.11.t=6.322,P<0.05).The positive rate of EdU in the control group cells[(70.47±10.42)%]was significantly higher than that in the hnRNPL KD group cel

关 键 词：核不均一核糖核蛋白L 口腔鳞状细胞癌 增殖 迁移 侵袭 

分 类 号：R739.8[医药卫生—肿瘤]