siRNA沉默X连锁凋亡抑制蛋白基因逆转人耐紫杉醇卵巢癌细胞耐药性的体内研究  

In vivo Study of siRNA Silencing XIAP Gene to Reverse Taxol-resistance in Human Ovarian Cancer Cells

作　　者：李冉红 岳驰 魏宝宝 刘娟 刘辉[2,3] LI Ranhong;YUE Chi;WEI Bao-bao;LIU Juan;LIU Hui(Department of Obstetrics and Gynecology,Henan Provincial People's Hospital,Zhengzhou 450003,China;Department of Obstetrics and Gynecology,West China Second University Hospital,Sichuan University,Chengdu 610041,China;Key Laboratory of Obstetric and Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education(Sichuan University),Chengdu 610041,China)

机构地区：[1]河南省人民医院妇产科,郑州450000 [2]四川大学华西第二医院妇产科,成都610041 [3]出生缺陷与相关妇儿疾病教育部重点实验室(四川大学),成都610041

出　　处：《四川大学学报（医学版）》2020年第3期320-324,共5页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技支撑计划项目(No.2013FZ0058)资助。

摘　　要：目的探究小干扰RNA(siRNA)沉默X连锁凋亡抑制蛋白(XIAP)基因表达对人耐紫杉醇卵巢癌细胞耐药性的逆转。方法36只BALB/C-nu雌性裸鼠皮下接种人耐紫杉醇卵巢癌细胞株A2780/T,建立耐紫杉醇卵巢癌裸鼠移植瘤模型,再随机分为6组(每组6只),A组:生理盐水组;B组:紫杉醇组;C组:siRNA-阴性对照(siRNA-NC)+生理盐水组;D组:siRNA-NC+紫杉醇组;E组:siRNA-XIAP+生理盐水组;F组:siRNA-XIAP+紫杉醇组。待接种处形成粟粒大小的移植瘤时,C^F组于瘤体处注射siRNA-XIAP或siRNA-NC,注射剂量为每次每只裸鼠10μg,每3 d注射1次,共注射9次。在D、F组第1次siRNA干扰后开始按裸鼠体质量给予2 mg/kg紫杉醇(C、E组注射生理盐水),每周1次,0.2 mL/次,腹腔内注射,共注射4次。A、B组不给予siRNA干扰,只进行紫杉醇或者生理盐水腹腔注射。siRNA处理27 d后处死裸鼠,取出瘤体测质量,计算抑瘤率;采用实时荧光定量PCR和Western blot分别检测各组移植瘤细胞中XIAP的基因和蛋白表达水平;TUNEL法检测各组移植瘤细胞的凋亡情况。结果转染阴性对照序列的生理盐水治疗(C组)抑瘤率最低、肿瘤质量最高、治疗效果最差,仅使用紫杉醇治疗(B组)、仅转染阴性对照序列的紫杉醇治疗组(D组)、仅沉默XIAP而不使用紫杉醇治疗(E组)三者效果相当,沉默XIAP且使用紫杉醇治疗组(F组)抑瘤率最高,治疗效果最好(P<0.05)。沉默XIAP后,F组与E组XIAP的基因和蛋白表达低于A^D组(P<0.05)。细胞凋亡指数:F组最高,A组及C组最低(P<0.05)。结论XIAP的沉默可逆转人耐紫杉醇卵巢癌细胞的耐药,增强紫杉醇的治疗效果。Objective To study the relationship between down-regulated expression of X linked inhibitor of apoptosis protein(XIAP)gene and the reversal effect of taxol-resistance by using siRNA interference technology in the taxol-resistant ovarian cancer.Methods Randomly assigned the nude mice into six groups(6 in each group).Group A:normal saline;Group B:taxol;Group C:siRNA-NC+normal saline;Group D:siRNA-NC+taxol;Group E:siRNA XIAP+normal saline;Group F:siRNA XIAP+taxol.Each group was dealt with the corresponding processing depending on the agreed protocol and the transplanted tumors had a multi-point injection with reagents related siRNA,one time every 3 days,9 times(27 d)in total.Taxol(2 mg/kg)was used in the intraperitoneal injection,0.2 mL every time,once a week,for four weeks.After 27 d of siRNA treatment,xenograft volumes and qualities were measured and the inhibitory rate was calculated;RNA expression levels and protein levels of XIAP gene in xenografts were detected respectively by real-time fluorescent quantitative PCR and Western blot.Apoptosis of the transplanted tumor cells was examined by TUNEL method.Results Among the six groups,the proliferation of transplanted tumor in Group F was the slowest,and the tumor inhibition rate was the highest compared with control Group A,followed by Group E,and the tumor inhibition rate was the lowest in Group C.Group F and E expressed the lowest XIAP mRNA and protein expressions(P<0.05,vs.the other 4 groups).The apoptosis rate was highest in Group F,followed by Group E,and lowest in Group A and C(P<0.05).Conclusion XIAP siRNA has synergy with taxol in taxol-resistant ovarian cancer cells.

关 键 词：XIAP基因 移植瘤 卵巢癌 siRNA 紫杉醇耐药 

分 类 号：R737.31[医药卫生—肿瘤]