长链非编码RNA ITGB1调控胆囊癌细胞耐药的分子机制  

Molecular mechanism of long non-coding RNA ITGB1 regulating drug resistance in gallbladder cancer cells

作　　者：厉冰[1] 尚华[2] 梅孝臣 门中俊 翟润 孙万日[1] Li Bing;Shang Hua;Mei Xiaochen;Men Zhongjun;Zhai Run;Sun Wanri(Department of General Surgery,Nanyang Central Hospital,Nanyang Central Hospital Affiliated to Henan University,Henan 473000,China;Nanyang Central Hospital,Department of Pediatric Digestive and Cardiovascular/Department of Pediatric Digestive and Cardiovascular,Nanyang Central Hospital Affiliated to Henan University,Nanyang 473000,China)

机构地区：[1]南阳市中心医院普外科,河南大学附属南阳市中心医院普外科,473000 [2]南阳市中心医院儿童消化心血管科/河南大学附属南阳市中心医院儿童消化心血管科,473000

出　　处：《中华实验外科杂志》2022年第11期2130-2132,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨长链非编码RNA(lncRNA)ITGB1在胆囊癌中表达水平及其对胆囊癌细胞化疗耐药的影响及分子机制。方法选取2018年1月到2021年1月南阳市中心医院收治的61例胆囊癌组织和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析胆囊癌组织和癌旁组织中lncRNA ITGB1表达水平。在人胆囊癌细胞株GBC-SD细胞采用浓度梯度递增法建立5-氟尿嘧啶(5-Fu)耐药细胞株GBC-SD/5-Fu,采用对照短发卡RNA(shRNA)和lncRNA ITGB1 shRNA慢病毒感染GBC-SD/5-Fu细胞系,建立lncRNA对照组和ITGB1 KD组。采用5-Fu处理lncRNA对照组和ITGB1 KD组细胞24 h后,采用细胞计数试剂盒(CCK-8)分析细胞增殖能力;采用体外移植瘤分析胆囊癌化疗耐药;采用流式细胞术分析5-Fu对细胞凋亡的影响;采用蛋白质印迹法(Western blot)分析两组细胞Atg5蛋白表达水平。组间比较采用t检验。结果癌旁组织lncRNA ITGB1表达水平(0.98±0.15)明显低于胆囊癌组织中(1.51±0.44),差异有统计学意义(t=9.054,P<0.05)。lncRNA对照组细胞48 h吸光度值(2.03±0.11)明显高于ITGB1 KD组(1.26±0.17),差异有统计学意义(t=9.450,P<0.05)。lncRNA对照组细胞体内增殖体积[(859.10±83.80)mm^(3)]明显高于ITGB1 KD组[(577.18±50.37)mm^(3)],差异有统计学意义(t=7.063,P<0.05)。lncRNA对照组细胞凋亡比例[(5.20±1.32)%]明显高于ITGB1 KD组[(33.62±3.88)%],差异有统计学意义(t=16.960,P<0.05)。lncRNA对照组细胞Atg5蛋白表达水平(1.14±0.07)明显高于ITGB1 KD组(0.52±0.12),差异有统计学意义(t=10.420,P<0.05)。结论lncRNA ITGB1在胆囊癌中呈高表达,参与胆囊癌细胞对5-Fu耐药过程,可能与自噬相关蛋白Atg5表达相关。Objective To investigate the expression level of long non-coding RNA(lncRNA)ITGB1 in gallbladder carcinoma and its effect on chemoresistance of gallbladder carcinoma cells and its molecular mechanism.Methods 61 cases of gallbladder cancer and adjacent tissues treated in our hospital from January 2018 to January 2021 were selected as research objects,and the expression level of lncRNA ITGB1 in gallbladder cancer and adjacent tissues was analyzed by fluorescent quantitative polymerase chain reaction(PCR).The 5-fluorouracil(5-Fu)resistant cell line GBC-SD/5-Fu was established in the human gallbladder cancer cell line GBC-SD by using the concentration gradient method.The GBC-SD/5-Fu cell line was infected with control short hairpin RNA(shRNA)and lncRNA ITGB1 shRNA lentivirus,and the lncRNA control group and ITGB1 KD group were established.After the cells of lncRNA control group and ITGB1KD group were treated with 5-Fu for 24 hours,the cell proliferation ability was analyzed with cell counting kit-8(CCK-8);The chemoresistance of gallbladder carcinoma was analyzed by transplantation in vitro.The effect of 5-Fu on apoptosis was analyzed by flow cytometry.The expression level of Atg5 protein was analyzed by Western blotting.The measurement data between groups were compared by t test.Results The expression level of lncRNA ITGB1 in adjacent tissues(0.98±0.15)was significantly lower than that in gallbladder carcinoma(1.51±0.44,t=9.054,P<0.05).The 48 h absorbance value of cells in the lncRNA control group(2.03±0.11)was significantly higher than that in the ITGB1 KD group(1.26±0.17,t=9.450,P<0.05).The proliferation volume of cells in lncRNA control group[(859.10±83.80)mm^(3)]was significantly higher than that of cells in ITGB1 KD group[(577.18±50.37)mm^(3),t=7.063,P<0.05].The percentage of apoptosis in lncRNA control group[(5.20±1.32)%]was significantly higher than that in ITGB1 KD group[(33.62±3.88)%,t=16.960,P<0.05].The expression level of Atg5 protein in lncRNA control group cells(1.14±0.07)was significantly highe

关 键 词：长链非编码RNA ITGB1 5-氟尿嘧啶 胆囊癌 耐药 自噬 

分 类 号：R735.8[医药卫生—肿瘤]