ADAM17小干扰RNA联合阿霉素抑制人前列腺癌小鼠移植瘤生长的实验研究  

Inhibitory Effect of ADAM17 Small Interfering RNA Combined with Adriamycin on the Growth of Transplanted Human Prostate Cancer in Mice

作　　者：李雪[1] 冯树强[1] 李然伟[1] 田文杰[1] 战鹏[1] Li Xue;Feng Shuqiang;Li Ranwei;Tian Wenjie;Zhan Peng(Department of Urology,the Second Hospital Affiliated to Jilin University,Changchun 130041,China)

机构地区：[1]吉林大学第二医院泌尿外科,长春130041

出　　处：《国际老年医学杂志》2020年第4期201-204,248,共5页International Journal of Geriatrics

基　　金：吉林省科学技术厅择优支持项目(20191102012YY)。

摘　　要：目的探讨小干扰RNA沉默ADAM17(siADAM17)基因表达联合化疗药阿霉素(ADM)对人前列腺癌PC3细胞株和小鼠皮下移植瘤生长的抑制作用。方法人前列腺癌PC3细胞分为5组,分别为:空白对照组、阴性对照组(转染空载体PGC-CONT)、siADAM17组(转染PGC-siADAM17)、ADM组(转染空载体PGC-CONT,0.05 g/L ADM,处理48 h)、联合应用组(转染PGC-siADAM17,0.05 g/L ADM,处理48 h)。噻唑蓝(MTT)法和流式细胞术(FCM)检测PC3细胞增殖和凋亡情况。利用上述5组PC3细胞悬液构建小鼠动物模型,分组及命名同PC3细胞,空白对照组注射PC3细胞;阴性对照组注射PC3/PGC-CONT细胞;siADAM17组注射PC3/PGC-siADAM17细胞;ADM组注射PC3/PGC-CONT细胞,同时给予ADM 1 mg/kg腹腔注射,1次/2d;联合应用组注射PC3/PGC-siADAM17细胞,同时给予ADM 1 mg/kg腹腔注射,1次/2d。原位末端标记(TUNEL)法检测小鼠移植瘤中细胞凋亡情况;SABC免疫组化检测瘤细胞增殖指数。结果MTT法显示siADAM17组、ADM组和联合应用组PC3细胞的生存率均明显低于阴性对照组,联合应用组明显低于siADAM17(P<0.0001)和ADM组(P<0.05)。FCM分析显示siADAM17组、ADM组和联合应用组细胞凋亡率均明显高于阴性对照组,而且联合应用组的凋亡率明显高于siADAM17组和ADM组(P<0.05)。siADAM17组、ADM组和联合应用组的肿瘤体积分别从第20 d、24 d和16 d开始明显小于阴性对照组(P<0.05);从第16 d起,联合应用组小鼠的肿瘤体积明显小于ADM组(P<0.05)和siADAM17组(P<0.001)。siADAM17组、ADM组和联合应用组TOPOⅡa增殖指数较阴性对照组均明显下降(P<0.001)。siADAM17组、ADM组和联合应用组移植瘤细胞的凋亡率较阴性对照组均明显增加(P<0.001),联合应用组移植瘤细胞的凋亡率明显高于siADAM17组和ADM组(P<0.05)。结论siADAM17或ADM单独应用均可抑制前列腺癌细胞增殖并诱导细胞凋亡,二者联合应用疗效更佳。Objective To investigate the inhibitory effect of siRNA silencing ADAM17(siADAM17)gene expression combined with adriamycin(ADM)on the growth of human prostate cancer PC3 cell line and subcutaneous transplanted tumor in mice.Methods PC3 cells were divided into 5 groups:blank control group,negative control group(transfected with empty vector PGC-CONT),siADAM17 group(transfected with PGC-siADAM17),ADM group(transfected with empty vector PGC-CONT and treated with 0.05 g/L of ADM for 48 h),combination group(transfected with PGC-siADAM17and treated with 0.05 g/L of ADM for 48 h).MTT and FCM were used to detect the proliferation and apoptosis of PC3 cells.PC3 cell suspension from the above five groups were subcutaneously injected into the mice.The mice in control group were injected with PC3 cells.The mice in negative control group were injected with PC3/PGC-CONT cells.The mice in siADAM17 group were injected with PC3/PGC-siADAM17 cells.The mice in ADM group were injected with PC3/PGC-CONT cells and ADM(1 mg/kg,intraperitoneally,once every 2 days).The mice in combination group were injected with PC3/PGC-siADAM17 cells and ADM(1 mg/kg,intraperitoneally,once every 2 days).TUNEL was used to examine the apoptosis of tumor cells in transplanted tumor.SABC immunohistochemical staining was used to examine the proliferation of tumor cell in transplanted tumor.Results MTT assay showed that the survival rate of PC3 cells in siADAM17 group,ADM group and combination group was significantly lower than that in negative control group respectively,the survival rate of PC3 cells in combination group was significantly lower than that in siADAM17(P<0.0001)and ADM group respectively(P<0.05).FCM analysis showed that the apoptosis rate in siADAM17 group,ADM group and combination group was significantly higher than that in negative control group respectively,and the apoptosis rate in combination group was significantly higher than that in siADAM17 group and ADM group respectively(P<0.05).The tumor volume in siADAM17 group,ADM group and combin

关 键 词：前列腺肿瘤 PC3细胞 siADAM17 ADM 动物模型 小鼠 

分 类 号：R737.25[医药卫生—肿瘤]