CAJ | 학술논문

目的探讨慢病毒介导shRNA沉默STAT3表达对结直肠癌生长的影响.方法分别用慢病毒干扰质粒pRNAT-shSTAT3、慢病毒空质粒pRNAT-GFP和慢病毒包装质粒混合物共转染293T细胞,获得慢病毒分别感染结直肠癌HT-29细胞,筛选培养后获得HT-29-shSTAT3、HT-29-GFP细胞株.将3种细胞株用于建立裸鼠皮下移植瘤模型,分为3组:第1组为HT-29组,第2组为HT-29-GFP组,第3组为HT-29-shSTAT3组,每组5只.MTT法检测细胞生长情况,流式细胞仪检测细胞周期变化,取瘤组织切片行CD34免疫组织化学观察肿瘤微血管密度(MVD)的变化.应用单因素方差分析检测结果.结果结直肠癌HT-29、HT-29-GFP细胞株生长能力明显强于HT-29-shSTAT3细胞株,3者比较,差异有统计学意义(F=632.50,P<0.05).HT-29-shSTAT3细胞株生长明显减慢,G0/G1期细胞占68.7%±2.9%,HT-29-GFP细胞株为38.5%±1.6%,HT-29细胞株为38.7%±2.3%;3者比较,差异有统计学意义(F=166.53,P<0.05).HT-29组、HT-29-GFP组和HT-29-sh STAT3组MVD分别为29±5、28±4、10±3,3组比较,差异有统计学意义(F=31.60,P<0.05).结论慢病毒介导的shRNA沉默STAT3表达能明显抑制结直肠癌的生长.
목적 탐토만병독개도shRNA침묵STAT3표체대결직장암생장적영향.방법 분별용만병독간우질립pRNAT-shSTAT3、만병독공질립pRNAT-GFP화만병독포장질립혼합물공전염293T세포,획득만병독분별감염결직장암HT-29세포,사선배양후획득HT-29-shSTAT3、HT-29-GFP세포주.장3충세포주용우건립라서피하이식류모형,분위3조:제1조위HT-29조,제2조위HT-29-GFP조,제3조위HT-29-shSTAT3조,매조5지.MTT법검측세포생장정황,류식세포의검측세포주기변화,취류조직절편행CD34면역조직화학관찰종류미혈관밀도(MVD)적변화.응용단인소방차분석검측결과.결과 결직장암HT-29、HT-29-GFP세포주생장능력명현강우HT-29-shSTAT3세포주,3자비교,차이유통계학의의(F=632.50,P<0.05).HT-29-shSTAT3세포주생장명현감만,G0/G1기세포점68.7%±2.9%,HT-29-GFP세포주위38.5%±1.6%,HT-29세포주위38.7%±2.3%;3자비교,차이유통계학의의(F=166.53,P<0.05).HT-29조、HT-29-GFP조화HT-29-sh STAT3조MVD분별위29±5、28±4、10±3,3조비교,차이유통계학의의(F=31.60,P<0.05).결론 만병독개도적shRNA침묵STAT3표체능명현억제결직장암적생장.
Objective To determine the effect of signal transducers and activators of transcription 3 (STAT3) gene silencing by shRNA mediated by lentiviral vector for the treatment of colorectal cancer. Methods The recombinant lentiviral vector pRNAT-shSTAT3, empty lentiviral vector pRNAT-GFP, and lentiviral packaging plasmids in supernatant were collected to transfect HT-29 cells for harvesting the HT-29-shSTAT3 cells and HT29-GFP cells. Fifteen male rats were divided into three groups (n = 5 ), and then they were inoculated with HT-29cells, HT-29-GFP cells and HT-29-shSTAT3 cells, respectively. Cell growth was assessed by MTT assay and the changes in cell cycle were detected by flow cytometry. The changes in microvessel density (MVD) of tumors were detected by immunohistochemistry. All data were analysed by one-way analysis of variance. Results The growth of HT-29-shSTAT3 cells was significantly suppressed compared with HT-29 and HT-29-GFP cells (F = 632.50,P ＜ 0. 05 ). The proportions of cells at the G0/G1 phase were 68.7% ± 2.9% in HT-29-shSTAT3 cells, 38.5% ±1.6% in HT-29-GFP cells and 38.7% ± 2.3% in HT-29 cells, with a significant difference among the three groups (F = 166.53, P ＜ 0.05 ). The MVDs of HT-29 cells, HT-29-GFP cells and HT-29-shSTAT3 cells were 29 ±5, 28 ±4 and 10 ±3, respectively, with a significant difference among the three groups (F=31.60, P ＜0.05). Conclusion STAT3 gene silencing by shRNA mediated by lentiviral vector can significantly inhibit the growth of colorectal cancer cells.