Musashi1对胃癌细胞增殖、转移和耐药的影响
Effect of Musashi1 on proliferation,metastasis and drug resistance of gastric cancer cells
马红岩 1崔玉凤 1蔡锋 2牛秉轩3
作者信息
- 1. 商丘医学高等专科学校,商丘 476100
- 2. 商丘市第一人民医院胃肠肝胆外科,商丘 476100
- 3. 新乡医学院,新乡 453000
- 折叠
摘要
目的 探讨RNA结合蛋白Musashi1在胃癌组织中的表达及对胃癌细胞的增殖、转移和耐药的影响.方法 选取2021年1月至2023年1月商丘市第一人民医院收治的74例手术切除的胃癌样本作为研究对象,并取癌旁组织作为对照.采用蛋白质印迹法(Western blot)分析癌旁组织和胃癌组织Musashi1蛋白表达水平;培养人胃癌细胞系HSC-38,采用Musashi1短发卡RNA(shRNA)慢病毒和对照慢病毒感染HSC-38,建立Musashi1低表达和对照细胞系,分别为Musashi1 KD组和对照组.采用细胞计数试剂盒(CCK-8)和克隆形成实验分析两组细胞的增殖能力;采用划痕实验和Transwell实验分析两组细胞的迁移和侵袭能力;采用Western blot分析两组细胞上皮间质转化能力;采用流式细胞术分析Musashi1对顺铂耐药细胞系的影响,组间比较采用t检验.结果 癌旁组织Musashi1蛋白表达水平(1.03±0.13)明显低于胃癌组织(1.87±0.29),差异有统计学意义(t=22.880,P<0.05).对照组细胞24 h吸光度值(2.06±0.14)明显高于Musashi1 KD组(1.41± 0.11),差异有统计学意义(t=8.958,P<0.05).对照组细胞克隆形成数量[(102.33±12.71)个]明显高于Musashi1 KD组[(63.83±6.67)个],差异有统计学意义(t=6.570,P<0.05).对照组细胞划痕愈合率[(83.50±8.22)%]明显高于Musashi1 KD组[(60.67±9.57)%],差异有统计学意义(t=4.141,P<0.05).对照组细胞侵袭数量[(121.83±10.80)个]明显高于Musashi1 KD组[(79.50±7.23)个],差异有统计学意义(t=7.980,P<0.05).对照组细胞上皮细胞E-钙黏蛋白(E-cadherin)蛋白表达水平(0.83±0.06)明显低于Musashi1 KD组(1.26±0.12),差异有统计学意义(t=3.147,P<0.05).对照组细胞上皮细胞N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达水平(0.85±0.10、1.05±0.13)明显高于 Musashi1 KD 组(0.48±0.08、0.69±0.08),差异有统计学意义(t=7.047、5.653,P<0.05).耐药对照组细胞凋亡水平[(33.67±6.31)%]明显低于耐药 Musashi1 KD 组[(77.67±5.57)%],差异有统计学意义(t=12.800,P<0.05).结论 RNA结合蛋白Musashi1在胃癌组织呈过表达,参与胃癌细胞的增殖、转移和耐药等恶性细胞生物学行为.
Abstract
Objective To investigate the expression of RNA binding protein Musashi1 in gastric cancer tissue and its effects on the proliferation,metastasis and drug resistance of gastric cancer cells.Methods A total of 74 surgically resected gastric cancer samples admitted to our hospital from January 2021 to January 2023 were selected as the research subjects,and adjacent cancerous tissues were taken as controls.The expression levels of Musashi1 protein in adjacent and gastric cancer tissues were detected by Western blotting.The human gastric cancer cell line HSC-38 was cultured,and infect with Musashi1 short hairpin RNA(shRNA)lentivirus and control lentivirus to establish Musashi1 low expression and control cell lines,serving as Musashi1 KD group and control group,respectively.The proliferation ability of two groups was analyzed using cell counting kit-8(CCK-8)and clone formation experiments.The migration and invasion abilities of the two groups were analyzed by wound healing and Transwell assays.The epithelial mesenchymal transition ability of two groups were analyzed by Western blotting.The effect of Musashi1 on cisplatin resistant cell lines was analyzed by flow cytometry.The comparison of inter group measurement data was done by t-test.Results The expression level of Musashi1 protein in adjacent cancerous tissue(1.03±0.13)was significantly lower than that in gastric cancer tissue(1.87±0.29,t=22.880,P<0.05).The 24-h absorbance values of the control group cells(2.06±0.14)was significantly higher than those of the Musashi1 KD group cells(1.41±0.11,t=8.958,P<0.05).The number of cell clones formed in the control group(102.33±12.71)was significantly greater than that in the Musashi1 KD group(63.83±6.67,t=6.570,P<0.05).The scratch healing rate in the control group[(83.50±8.22)%]was significantly higher than that in the Musashi1 KD group[(60.67±9.57)%,t=4.141,P<0.05].The number of invasive cells in the control group[(121.83±10.80)]was significantly greater than that in the Musashi1 KD group[(79.50±7.23),t=7.980,P<0.05].The expression level of E-Cadherin protein in the epithelial cells of the control group(0.83±0.06)was significantly lower than that of the Musashi1 KD group(1.26±0.12,t=3.147,P<0.05).The expression levels of N-cadherin and Vimentin proteins in the epithelial cells of the control group(0.85±0.10,1.05±0.13)were significantly higher than those in the Musashi1 KD group(0.48±0.08,0.69±0.08,t=7.047,5.653,P<0.05).The apoptosis rate of cells in the drug-resistant control group[33.67±6.31)%]was significantly lower than that in the drug-resistant Musashi1 KD group[(77.67±5.57)%,t=12.800,P<0.05].Conclusion RNA binding protein Musashi1 in gastric cancer tissue significantly increased,which is involved in malignant cell biology behav-iors such as proliferation,metastasis,and drug resistance of gastric cancer cells.
关键词
RNA结合蛋白Musashi1/胃癌/增殖/转移/耐药Key words
RNA binding protein Musashi1/Gastric cancer/Proliferation/Transfer/Drug resistance引用本文复制引用
出版年
2024