中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
10期
1422-1424,后插二
,共4页
申培红%樊青霞%张红新%张威%张云汉
申培紅%樊青霞%張紅新%張威%張雲漢
신배홍%번청하%장홍신%장위%장운한
BCSG1%RNA干扰%转染%抑制
BCSG1%RNA榦擾%轉染%抑製
BCSG1%RNA간우%전염%억제
BCSG1%RNA-interference%Transfection%Inhibition
目的 观察RNA干扰(RNAi)对MCF7细胞株BCSG-1基因表达的抑制作用.方法 针对BCSG-1的已知cDNA序列,设计并体外转录合成4条特异性siRNA(BCSG-1-siRNA-1、BCSG-1-siRNA-2、BCSG-1-siRNA-3、BCSG-1-siRNA-4),1条非特异性siRNA,用脂质体介导转染MCF-7细胞48 h,同时设空白对照组(空脂质体)、正常MCF7对照组(不转染).采用免疫细胞化学及半定量逆转录-聚合酶链反应(RT-PCR)检测7组细胞中BCSG-1蛋白及mRNA的表达,免疫细胞化学7组中BCSG-1蛋白表达分别为4.27±0.12、4.19±0.22、4.15±0.14、4.17±0.13、7.92±0.22、8.02±0.13、8.02±0.13;RT-PCR测7组细胞中BCSG-1 mRNA的表达分别为0.624±0.010、0.626±0.013、0.634±0.008、0.631±0.010、0.976±0.076、0.983±0.052、1.014±0.034.结果 7组中BCSG-1蛋白表达结果比较,差异有统计学意义(P＜0.01);mRNA的表达结果比较,差异有统计学意义(P＜0.01).与各对照组比较,BCSG-1-siRNA转染的4组细胞中BCSG-1蛋白及mRNA的表达均减弱,差异均有统计学意义(P＜0.05).结论 BCSG-1-siRNA能有效抑制乳腺癌MCF-7细胞中BCSG-1蛋白及mRNA的表达.
目的 觀察RNA榦擾(RNAi)對MCF7細胞株BCSG-1基因錶達的抑製作用.方法 針對BCSG-1的已知cDNA序列,設計併體外轉錄閤成4條特異性siRNA(BCSG-1-siRNA-1、BCSG-1-siRNA-2、BCSG-1-siRNA-3、BCSG-1-siRNA-4),1條非特異性siRNA,用脂質體介導轉染MCF-7細胞48 h,同時設空白對照組(空脂質體)、正常MCF7對照組(不轉染).採用免疫細胞化學及半定量逆轉錄-聚閤酶鏈反應(RT-PCR)檢測7組細胞中BCSG-1蛋白及mRNA的錶達,免疫細胞化學7組中BCSG-1蛋白錶達分彆為4.27±0.12、4.19±0.22、4.15±0.14、4.17±0.13、7.92±0.22、8.02±0.13、8.02±0.13;RT-PCR測7組細胞中BCSG-1 mRNA的錶達分彆為0.624±0.010、0.626±0.013、0.634±0.008、0.631±0.010、0.976±0.076、0.983±0.052、1.014±0.034.結果 7組中BCSG-1蛋白錶達結果比較,差異有統計學意義(P＜0.01);mRNA的錶達結果比較,差異有統計學意義(P＜0.01).與各對照組比較,BCSG-1-siRNA轉染的4組細胞中BCSG-1蛋白及mRNA的錶達均減弱,差異均有統計學意義(P＜0.05).結論 BCSG-1-siRNA能有效抑製乳腺癌MCF-7細胞中BCSG-1蛋白及mRNA的錶達.
목적 관찰RNA간우(RNAi)대MCF7세포주BCSG-1기인표체적억제작용.방법 침대BCSG-1적이지cDNA서렬,설계병체외전록합성4조특이성siRNA(BCSG-1-siRNA-1、BCSG-1-siRNA-2、BCSG-1-siRNA-3、BCSG-1-siRNA-4),1조비특이성siRNA,용지질체개도전염MCF-7세포48 h,동시설공백대조조(공지질체)、정상MCF7대조조(불전염).채용면역세포화학급반정량역전록-취합매련반응(RT-PCR)검측7조세포중BCSG-1단백급mRNA적표체,면역세포화학7조중BCSG-1단백표체분별위4.27±0.12、4.19±0.22、4.15±0.14、4.17±0.13、7.92±0.22、8.02±0.13、8.02±0.13;RT-PCR측7조세포중BCSG-1 mRNA적표체분별위0.624±0.010、0.626±0.013、0.634±0.008、0.631±0.010、0.976±0.076、0.983±0.052、1.014±0.034.결과 7조중BCSG-1단백표체결과비교,차이유통계학의의(P＜0.01);mRNA적표체결과비교,차이유통계학의의(P＜0.01).여각대조조비교,BCSG-1-siRNA전염적4조세포중BCSG-1단백급mRNA적표체균감약,차이균유통계학의의(P＜0.05).결론 BCSG-1-siRNA능유효억제유선암MCF-7세포중BCSG-1단백급mRNA적표체.
Objective To investigate the suppressive effects of siRNA on the BCSG-1 gene expression in breast cancer MCF-7 cells. Methods The four-pair designed and chemically synthesized in vitro of small interfering RNA (siRNA) sequences ( BCSG-1-siRNA-1 ,-2,-3 and 4) for BCSG-1, and one pair of non-specific siRNA sequence were transducted into plasmid vectors, following by transfection into into MCF-7 cells. Blank control group (empty vector transfection) and normal control group (no transfection) were set up. After 48 h, the mRNA and protein expression levels of BCSG-1 in transfected cells were detected by using reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry respectively. Results The expression of BCSG1 protein was 4.27 ±0. 12, 4.19 ±0.22, 4.15 ± 0.14,4.17 ±0.13, 7.92 ±0.22, 8.02 ±0.13, 8.02 ±0.13, and that of BCSG1 mRNA was 0.624 ±0.010,0.626±0.013, 0.634 ±0.008, 0.631 ±0.010, 0.976±0.076, 0.983 ±0.052, 1.014±0.034 respectively. As compared with control groups, there was significant difference in the expression of BCSG-1 mRNA and protein among the above groups (P ＜0.01 ). The expression level of BCSG-1 mRNA and protein in the groups of MFC-1 cells transfected with BCSG-1-siRNA was lower than that in control groups.Conclusion Expression of BCSG-1 gene can be effectively inhibited by siRNA in MCF7 cells.
