胃癌组织中miR-45表达及其对胃癌原代培养细胞5-FU敏感性的影响  

Expression of miR-45 in gastric cancer tissue and its effect on sensitivity of primary cultured gastric cancer cells to 5-fluorouracil

作　　者：檀碧波[1] 张明月 赵群[1] 范立侨[1] 刘庆伟[1] 李勇[1] TAN Bi-bo;ZHANG Ming-yue;ZHAO Qun;FAN Li-qiao;LIU Qing-wei;LI Yong(Third Department of Surgery,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,P.R.China)

机构地区：[1]河北医科大学第四医院外三科,河北石家庄050011

出　　处：《中华肿瘤防治杂志》2020年第14期1133-1137,共5页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的胃癌中微小RNA-145(miR-145)存在异常表达,但miR-145与胃癌化疗耐药的关系罕见报道。本研究探讨胃癌组织中miR-145表达与肿瘤细胞对氟尿嘧啶(5-fluorouracil,5-FU)化疗敏感性关系及临床意义,为改善5-FU对胃癌的化疗效果提供依据。方法收集2017-06-01-2019-10-31河北医科大学第四医院外科确诊并手术的70例胃癌患者肿瘤及癌旁组织(距肿瘤边缘>3cm,镜下未见到癌或不典型增生组织),实时荧光定量逆转录-聚合酶链反应(real-time quantitative reverse transcription polymerase chain reaction,qRT-PCR)法检测组织中miR-145及多药耐药基因1(multidrug resistance gene 1,MDR1)、B淋巴细胞/白血病-2(B lymphatic leukaemia 2,Bcl-2)、Livin和胸苷酸合成酶(thymidylate synthase,TS)基因的mRNA表达;免疫组化法检测P-糖蛋白(P-glycoprotein,P-gp)、Bcl-2、Livin和TS蛋白表达;噻唑蓝(thiazolyl blue,MTT)法检测胃癌细胞对5-FU的体外敏感性。结果miR-145在胃癌组织水平(0.389 0±0.111 8)低于癌旁正常组织(0.505 7±0.202 0),差异有统计学意义,t=-4.229,P<0.001;胃癌组织MDR1、Bcl-2和TS的mRNA水平均高于癌旁正常组织,均P<0.05。MTT结果显示,肿瘤组织细胞在5-FU作用后活性为(46.081±15.721)%;miR-145高水平的肿瘤细胞对5-FU的活性(42.223±13.478)%低于miR-145低水平者(50.406±17.090)%,t=-2.236,P=0.029;在胃癌组织中P-gp、Bcl-2和TS蛋白阳性的肿瘤组织对5-FU的活性高于阴性表达者(P<0.05),Livin表达与肿瘤组织对5-FU的活性无关(P>0.05)。miR-145与Bcl-2(r=-0.510 6,P<0.001)、TS(r=-0.403 2,P=0.000 5)的mRNA表达关系经Spearman秩相关分析发现均存在中度负相关,与MDR1(r=-0.203 2,P=0.091 6)、Livin(r=-0.174 3,P=0.149 0)的mRNA无统计学意义的相关。结论 miR-145可能通过调节Bcl-2、TS等耐药相关基因参与了胃癌对5-FU的耐药形成。OBJECTIVE It has been reported that microRNA-145(miR-145)is abnormally expressed in gastric cancer,but the relationship between miR-145 and chemotherapy resistance in gastric cancer keeps rare.Purpose of this study was to explore relationship between miR-145 in gastric cancer tissues and chemosensitivity of 5-fluorouracil(5-FU),and to analyze the clinical significance,and improve effect of 5-FU to gastric cancer.METHODS Specimens of gastric cancer tissues and cancer-adjacent normal tissues from 70 patients from 1 st,June,2017 to 31,October,2019 were collected.Expressions of miR-145 and mRNA of MDR1,Bcl-2,Livin,TSwere detected with real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR),and expressions of P-gp,Bcl-2,Livin,TS proteins were detected with immunohistochemistry(IHC).MTT assay was utilized to test sensitivity of gastric cancer cells to 5-FUin vitro.RESULTS Level of miR-145 in gastric cancer tissues(0.3890±0.1118)was much lower than that in cancer-adjacent normal tissues(0.5057±0.2020),t=-4.229,P<0.001;levels of MDR1,Bcl-2,TS mRNAs were all higher than in cancer-adjacent normal tissues(P<0.05).Results of MTT assay detected that relative cell activity was(46.081±15.721)%;cell activity in high-level miR-145 group(42.223±13.478)%was lower than that in low-level miR-145 group(50.406±17.090)%,t=-2.236,P=0.029;cell activity in positive expressed P-gp,Bcl-2,TS proteins groups were higher in negative expressed groups(P<0.05),while no association was verified between cell activity(treated with5-FU)and expression of Livin(P>0.05).Negative correlation was presented between miR-145 and Bcl-2,miR-145 and TS(r=-0.5106,P<0.001;r=-0.4032,P=0.0005),while no significant correlations were found between miR-145 and MDR1,miR-145 and Livin(r=-0.2032,P=0.0916;r=-0.1743,P=0.1490).CONCLUSION miR-145 may be involved in drug resistance of gastric cancer cells to 5-FU by regulating Bcl-2,TS genes.

关 键 词：微小RNA145 胃癌 氟尿嘧啶 药物抵抗 

分 类 号：R735.2[医药卫生—肿瘤]