中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2010年
11期
819-824
,共6页
杨增%严丽%张峰%闫庆辉%宋伟庆%王凤安%蔡建辉
楊增%嚴麗%張峰%閆慶輝%宋偉慶%王鳳安%蔡建輝
양증%엄려%장봉%염경휘%송위경%왕봉안%채건휘
乳腺肿瘤%RNA干扰%STAT3转录因子%MCF7细胞系%小鼠,裸
乳腺腫瘤%RNA榦擾%STAT3轉錄因子%MCF7細胞繫%小鼠,裸
유선종류%RNA간우%STAT3전록인자%MCF7세포계%소서,라
Breast neoplasms%RNA interference%STAT3 transcription factor%MCF7 cells line
目的 观察沉默信号转导和转录激活因子3(STAT3)基因对乳腺癌细胞MCF7体内外生长的抑制作用,探讨STAT3基因作为乳腺癌治疗靶点的可行性和有效性.方法 应用RNA干扰技术抑制MCF7细胞STAT3基因的表达,半定量逆转录聚合酶链反应(RT-PCR)和Western blot法检测STAT3 mRNA及蛋白的表达,流式细胞仪检测MCF7细胞的凋亡率,四甲基偶氮唑蓝(MTT)法检测MCF7细胞的增殖情况.在裸鼠皮下移植转染了STAT3 siRNA的MCF7细胞,观察其成瘤性.半定量RT-PCR和Western blot法检测成瘤标本的STAT3 mRNA和蛋白的表达.结果 转染48 h后,STAT3 siRNA转染组、非特异siRNA转染组和空白对照组的STAT3 mRNA表达量分别为0.327±0.020、1.035±0.050和1.093±0.018;转染72 h后,3组STAT3蛋白表达量分别为0.153±0.006、1.320±0.033和1.374±0.022,STAT3 siRNA转染组的STAT3 mRNA和蛋白表达水平,较非特异siRNA转染组和空白对照组均明显降低(P＜0.05).MTT实验结果显示,STAT3 siRNA转染组和非特异siRNA转染组的细胞生长抑制率分别为(44.00±5.10)%和(16.10±1.05)%,STAT3 siRNA转染组的细胞增殖能力明显下降(P＜0.05).流式细胞仪分析显示,STAT3 siRNA转染组、非特异siRNA转染组和空白对照组的细胞凋亡率分别为(14.79±0.22)%、(8.45±0.43)%和(7.06±0.71)%,STAT3 siRNA转染组明显高于非特异siRNA转染组和空白对照组(P＜0.05).STAT3siRNA转染组、非特异siRNA转染组和空白对照组在裸鼠体内最终的成瘤体积分别为(41.15±12.17)mm3、(101.36±21.90)mm3和(118.45±24.68)mm3,移植瘤重量分别为(21.4±10.6)mg、(57.2±21.9)mg和(88.6±12.2)mg,STAT3 siRNA转染组的移植瘤体积和重量明显低于非特异siRNA转染组和空白对照组(P＜0.05).STAT3 siRNA转染组移植瘤组织的STAT3 mRNA和蛋白表达水平均明显降低.结论 靶向STAT3的siRNA可以抑制乳腺癌MCF7细胞体内外的生长,STAT3有可能成为新的乳腺癌治疗靶点.
目的 觀察沉默信號轉導和轉錄激活因子3(STAT3)基因對乳腺癌細胞MCF7體內外生長的抑製作用,探討STAT3基因作為乳腺癌治療靶點的可行性和有效性.方法 應用RNA榦擾技術抑製MCF7細胞STAT3基因的錶達,半定量逆轉錄聚閤酶鏈反應(RT-PCR)和Western blot法檢測STAT3 mRNA及蛋白的錶達,流式細胞儀檢測MCF7細胞的凋亡率,四甲基偶氮唑藍(MTT)法檢測MCF7細胞的增殖情況.在裸鼠皮下移植轉染瞭STAT3 siRNA的MCF7細胞,觀察其成瘤性.半定量RT-PCR和Western blot法檢測成瘤標本的STAT3 mRNA和蛋白的錶達.結果 轉染48 h後,STAT3 siRNA轉染組、非特異siRNA轉染組和空白對照組的STAT3 mRNA錶達量分彆為0.327±0.020、1.035±0.050和1.093±0.018;轉染72 h後,3組STAT3蛋白錶達量分彆為0.153±0.006、1.320±0.033和1.374±0.022,STAT3 siRNA轉染組的STAT3 mRNA和蛋白錶達水平,較非特異siRNA轉染組和空白對照組均明顯降低(P＜0.05).MTT實驗結果顯示,STAT3 siRNA轉染組和非特異siRNA轉染組的細胞生長抑製率分彆為(44.00±5.10)%和(16.10±1.05)%,STAT3 siRNA轉染組的細胞增殖能力明顯下降(P＜0.05).流式細胞儀分析顯示,STAT3 siRNA轉染組、非特異siRNA轉染組和空白對照組的細胞凋亡率分彆為(14.79±0.22)%、(8.45±0.43)%和(7.06±0.71)%,STAT3 siRNA轉染組明顯高于非特異siRNA轉染組和空白對照組(P＜0.05).STAT3siRNA轉染組、非特異siRNA轉染組和空白對照組在裸鼠體內最終的成瘤體積分彆為(41.15±12.17)mm3、(101.36±21.90)mm3和(118.45±24.68)mm3,移植瘤重量分彆為(21.4±10.6)mg、(57.2±21.9)mg和(88.6±12.2)mg,STAT3 siRNA轉染組的移植瘤體積和重量明顯低于非特異siRNA轉染組和空白對照組(P＜0.05).STAT3 siRNA轉染組移植瘤組織的STAT3 mRNA和蛋白錶達水平均明顯降低.結論 靶嚮STAT3的siRNA可以抑製乳腺癌MCF7細胞體內外的生長,STAT3有可能成為新的乳腺癌治療靶點.
목적 관찰침묵신호전도화전록격활인자3(STAT3)기인대유선암세포MCF7체내외생장적억제작용,탐토STAT3기인작위유선암치료파점적가행성화유효성.방법 응용RNA간우기술억제MCF7세포STAT3기인적표체,반정량역전록취합매련반응(RT-PCR)화Western blot법검측STAT3 mRNA급단백적표체,류식세포의검측MCF7세포적조망솔,사갑기우담서람(MTT)법검측MCF7세포적증식정황.재라서피하이식전염료STAT3 siRNA적MCF7세포,관찰기성류성.반정량RT-PCR화Western blot법검측성류표본적STAT3 mRNA화단백적표체.결과 전염48 h후,STAT3 siRNA전염조、비특이siRNA전염조화공백대조조적STAT3 mRNA표체량분별위0.327±0.020、1.035±0.050화1.093±0.018;전염72 h후,3조STAT3단백표체량분별위0.153±0.006、1.320±0.033화1.374±0.022,STAT3 siRNA전염조적STAT3 mRNA화단백표체수평,교비특이siRNA전염조화공백대조조균명현강저(P＜0.05).MTT실험결과현시,STAT3 siRNA전염조화비특이siRNA전염조적세포생장억제솔분별위(44.00±5.10)%화(16.10±1.05)%,STAT3 siRNA전염조적세포증식능력명현하강(P＜0.05).류식세포의분석현시,STAT3 siRNA전염조、비특이siRNA전염조화공백대조조적세포조망솔분별위(14.79±0.22)%、(8.45±0.43)%화(7.06±0.71)%,STAT3 siRNA전염조명현고우비특이siRNA전염조화공백대조조(P＜0.05).STAT3siRNA전염조、비특이siRNA전염조화공백대조조재라서체내최종적성류체적분별위(41.15±12.17)mm3、(101.36±21.90)mm3화(118.45±24.68)mm3,이식류중량분별위(21.4±10.6)mg、(57.2±21.9)mg화(88.6±12.2)mg,STAT3 siRNA전염조적이식류체적화중량명현저우비특이siRNA전염조화공백대조조(P＜0.05).STAT3 siRNA전염조이식류조직적STAT3 mRNA화단백표체수평균명현강저.결론 파향STAT3적siRNA가이억제유선암MCF7세포체내외적생장,STAT3유가능성위신적유선암치료파점.
Objective To observe the effect of signal transducers and activators of transcription 3 (STAT3) gene silence on the growth of breast cancer cell line MCF7 in vitro and in vivo and discuss the feasibility and effectiveness of STAT3 used as gene therapeutic target for breast cancer. Methods Human breast cancer cell line MCF7 cells were divided into 3 groups:mock control group, control group transfected with scrambled sequence siRNA, and experimental group transfectod with STAT3 siRNA. The STAT3 mRNA and protein levels were detected by semi-quantity RT-PCR and Western blotting, respectively. The cell proliferation and apoptosis were examined by MTT method and flow cytometry. MCF7 cells treated with STAT3-siRNA were transplanted subcutaneously in nude mice and their tumorgenic ability was observed. The STAT3 mRNA and protein levels of the samples from nude mice of different groups were detected by semiquantity RT-PCR and Western blotting and compared. Results After treatment with STAT3-siRNA, STAT3 mRNA (0.327 ±0.020 vs. 1.035 ±0.050, 1.093 ±0.018) and ptotein (0. 153 ±0.006 vs. 1.320 ±0.033, 1. 374 ± 0. 022) levels in the MCF7 cells transfected with STAT3-siRNA were significantly lower than that in the two control groups ( P ＜ 0.05). MTT assay showed that after transfection of the STAT3-siRNA into MCF7 cells, cell proliferation was significantly reduced and the cell growth inhibition ratio in the STAT3-siRNA group was (44.00±5.10)%, significantly higher than that in the control group (16.1 ±1.05 )% (P ＜ 0. 05 ). Flow cytometry results suggested that more apoptosis was observed in the STAT3-siRNA group. The apoptosis rate was ( 14.79±0.22)%, much higher than that in the control group [(7.06 ±0.71 ) %, ( 8.45 ± 0.43 ) %, P ＜ 0.05]. The tumor growth in the experimental group was significantly slower than that in the two control groups. On the 22th day after transplantation, the tumor weight [(21.4 ±10.6) mg vs. (88.6±12.2) mg, (57.2 ±21.9) mg] and volume [(41.15 ±12.17) mm3 vs.( 118.45 ± 24.68) mm3, ( 101.36 ± 21.90) mm3] in the experimental group were significantly lower than that in the two control groups (P ＜ 0.05 ). The STAT3 mRNA and protein levels of the samples from nude mice in the experimental group were significantly lower than that in the two control groups. Conclusion siRNA targeting STAT3 can inhibit the proliferation of MCF7 cells in vitro and in vivo. STAT3 may become a novel therapeutic target for breast cancer.
