CAJ | 학술논문

目的观察携带靶向信号转导和转录活化因子3(STAT3)的小分子干扰RNA(siRNA)重组慢病毒(LV-STAT3 siRNA)与塞卡替尼(AZD0530)联合治疗在体内外抑制胶质瘤生长的效果.方法以人脑胶质瘤U87、U251细胞株和U87细胞BALB/c-nu裸鼠动物模型为研究对象,分为阴性对照组、空载体组、LV-STAT3 siRNA组、AZD0530组和LV-STAT3 siRNA与AZD0530联合治疗组.在体外实验中应用细胞计数试剂盒(CCK-8)方法检测细胞增殖,应用流式细胞仪检测细胞凋亡.在体内实验中,应用小动物活体成像技术动态监测肿瘤生长,3周后处死裸鼠,行免疫组织化学染色.结果体外实验中联合治疗组对U87和U251细胞增殖的平均抑制率分别达28.60％、36.36％(P＜0.05);在U87细胞中,联合治疗组凋亡率最高[(55.28±1.59)％],其次为LV-STAT3siRNA组和AZD0530组[(46.38±1.76)％、(35.39±1.23)％],3组之间差异有统计学意义(P＜0.05),在U251细胞中结果类似;体内实验中,第1周时LV-STAT3 siRNA组(8.05±1.09)、AZD0530组(7.54±1.07)和联合治疗组(7.19 ±0.73)发光值低于空载体组(11.14±1.59),差异有统计学意义(P＜0.05);第2周,AZD0530组(79.63 ±8.25)和空载体组(92.23±17.34)吸光度(A)值差异无统计学意义(P＞0.05),LV-STAT3 siRNA组(63.78 ±13.46)和联合治疗组(61.57 ±7.84)A值低于AZD0530组和空载体组,差异有统计学意义(P＜0.05).免疫组织化学显示,磷酸化STAT3和B细胞淋巴瘤/白血病-2(bcl-2)蛋白在阴性对照组、空载体组和AZD0530组的表达量分别约是LV-STAT3 siRNA组和联合治疗组的3、4倍(P＜0.05).结论体外实验中联合治疗产生明显的协同抑瘤作用;体内实验中LV-STAT3 siRNA抑瘤效果显著,AZD0530效果不理想.
목적 관찰휴대파향신호전도화전록활화인자3(STAT3)적소분자간우RNA(siRNA)중조만병독(LV-STAT3 siRNA)여새잡체니(AZD0530)연합치료재체내외억제효질류생장적효과.방법 이인뇌효질류U87、U251세포주화U87세포BALB/c-nu라서동물모형위연구대상,분위음성대조조、공재체조、LV-STAT3 siRNA조、AZD0530조화LV-STAT3 siRNA여AZD0530연합치료조.재체외실험중응용세포계수시제합(CCK-8)방법검측세포증식,응용류식세포의검측세포조망.재체내실험중,응용소동물활체성상기술동태감측종류생장,3주후처사라서,행면역조직화학염색.결과 체외실험중연합치료조대U87화U251세포증식적평균억제솔분별체28.60％、36.36％(P＜0.05);재U87세포중,연합치료조조망솔최고[(55.28±1.59)％],기차위LV-STAT3siRNA조화AZD0530조[(46.38±1.76)％、(35.39±1.23)％],3조지간차이유통계학의의(P＜0.05),재U251세포중결과유사;체내실험중,제1주시LV-STAT3 siRNA조(8.05±1.09)、AZD0530조(7.54±1.07)화연합치료조(7.19 ±0.73)발광치저우공재체조(11.14±1.59),차이유통계학의의(P＜0.05);제2주,AZD0530조(79.63 ±8.25)화공재체조(92.23±17.34)흡광도(A)치차이무통계학의의(P＞0.05),LV-STAT3 siRNA조(63.78 ±13.46)화연합치료조(61.57 ±7.84)A치저우AZD0530조화공재체조,차이유통계학의의(P＜0.05).면역조직화학현시,린산화STAT3화B세포림파류/백혈병-2(bcl-2)단백재음성대조조、공재체조화AZD0530조적표체량분별약시LV-STAT3 siRNA조화연합치료조적3、4배(P＜0.05).결론 체외실험중연합치료산생명현적협동억류작용;체내실험중LV-STAT3 siRNA억류효과현저,AZD0530효과불이상.
Objective To explore the anti-glioma effect of recombinant lentivirus carried small-interference RNA (siRNA) targeting signal transducers and activators of transcription 3 (STAT3) combined therapy with saracatinib (AZD0530) in vivo and in vitro.Methods Mice models were established by injecting transfected U87,U251 cells into BALB/c-nu nude mice.The mice were randomly divided into 5 groups:the negative control group,empty vector group,LV-STAT3 siRNA group,AZD0530 group and LV-STAT3 siRNA combined with AZD0530group.In vitro,Cell Counting Kit-8 (CCK-8) methods were applied to detect cell proliferation and apoptosis were detected by flow cytometry.The tumors were imaged every seven day in vivo.After three weeks treatment,immunohistochemical were performed.Results In vitro,combined treatment group demonstrate significant inhibition on U87 and U251 cell proliferation with the average rate of 28.60％,36.36％,respectively.(P ＜ 0.05) ; In U87 cells,apoptosis rate was the highest in the combined treatment group (55.28 ± 1.59) ％,followed by LV-STAT3 siRNA group and AZD0530 group [(46.38 ± 1.76)％,(35.39 ± 1.23)％],a statistically significant difference between three groups (P ＜ 0.05),U251 cells show similar results ; in vivo,the LV-STAT3 siRNA group (8.05 ± 1.09),AZD0530 group (7.54 ± 1.07) and combined treatment group (7.19 ± 0.73) emission value is lower than the empty vector group (11.14 ± 1.59) in first week,the difference is statistically significant (P ＜0.05) ; While AZD0530 group (79.63 ± 8.25) and empty vector group (92.23 ± 17.34) absorbance values were not statistically different (P ＞ 0.05),LV-STAT3 siRNA group (63.78 ± 13.46) and combined treatment group (61.57 ± 7.84) was lower than AZD0530 group and empty vector group in the second week,the difference was statistically significant (P ＜ 0.05).The results of Immunostaining Showed:Phosphorylated STAT3 and B cell lymphoma/lewkmia-2 (bcl-2) protein were expressed in the negative control group,empty vector group and AZD0530 group were about three times in LV-STAT3 siRNA group and four times in combined treatment group (P ＜ 0.05).Conclusion In vitro,LV-STAT3 siRNA combined with AZD0530 showed significantly inhibitory effect of tumor.in vivo,STAT3 siRNA was significantly inhibitory effect of tumor,the effect of AZD0530 was not ideal.