天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2014年
9期
863-866
,共4页
胰腺肿瘤%癌%基因沉默%细胞增殖%干扰素Ⅱ型%转染%半数抑制浓度%细胞因子信号转导蛋白抑制因子
胰腺腫瘤%癌%基因沉默%細胞增殖%榦擾素Ⅱ型%轉染%半數抑製濃度%細胞因子信號轉導蛋白抑製因子
이선종류%암%기인침묵%세포증식%간우소Ⅱ형%전염%반수억제농도%세포인자신호전도단백억제인자
pancreatic neoplasms%carcinoma%gene silencing%cell proliferation%interferon type II%transfection%IC50%suppressor of cytokine signaling proteins
目的：在人胰腺癌细胞中观察细胞因子信号转导抑制因子1（SOCS1）沉默对细胞增殖及干扰素-γ（IFN-γ）敏感性的影响，探讨SOCS1作为胰腺癌治疗靶点的可能。方法 Western blot及PCR验证SOCS1干扰序列沉默人胰腺癌细胞系PANC1中SOCS1的表达；给予IFN-γ刺激后，采用Western blotting方法观察转录激活因子（STAT）1及磷酸化STAT（pSTAT）1的变化；采用定量PCR的方法观察IFN-γ调节因子1（IRF-1）表达水平变化；MTT法检测胰腺癌细胞对IFN-γ敏感性的变化；细胞计数的方法观察细胞的增殖速度；采用流式细胞术的方法检测细胞周期的变化。结果将SOCS1干扰序列转染PANC1细胞后，SOCS1 mRNA及蛋白表达水平均明显下降。沉默SOCS1表达后，pshSOCS1-PANC1组细胞的IRF-1 mRNA水平及pSTAT1蛋白表达水平均显著升高（P<0.05），IFN-γ对PANC1细胞的半数抑制浓度（IC50）显著降低（P<0.01）；转染72 h后PANC1细胞数量较对照组显著减少（P<0.05）；SOCS1表达抑制后PANC1细胞G0/G1期细胞比例明显升高，而S期和G2/M期细胞比例明显减小，与对照组比较差异均有统计学意义。结论 SOCS1表达抑制后，人胰腺癌细胞株PANC1的增殖能力下降，并且对IFN-γ的敏感性增强。
目的：在人胰腺癌細胞中觀察細胞因子信號轉導抑製因子1（SOCS1）沉默對細胞增殖及榦擾素-γ（IFN-γ）敏感性的影響，探討SOCS1作為胰腺癌治療靶點的可能。方法 Western blot及PCR驗證SOCS1榦擾序列沉默人胰腺癌細胞繫PANC1中SOCS1的錶達；給予IFN-γ刺激後，採用Western blotting方法觀察轉錄激活因子（STAT）1及燐痠化STAT（pSTAT）1的變化；採用定量PCR的方法觀察IFN-γ調節因子1（IRF-1）錶達水平變化；MTT法檢測胰腺癌細胞對IFN-γ敏感性的變化；細胞計數的方法觀察細胞的增殖速度；採用流式細胞術的方法檢測細胞週期的變化。結果將SOCS1榦擾序列轉染PANC1細胞後，SOCS1 mRNA及蛋白錶達水平均明顯下降。沉默SOCS1錶達後，pshSOCS1-PANC1組細胞的IRF-1 mRNA水平及pSTAT1蛋白錶達水平均顯著升高（P<0.05），IFN-γ對PANC1細胞的半數抑製濃度（IC50）顯著降低（P<0.01）；轉染72 h後PANC1細胞數量較對照組顯著減少（P<0.05）；SOCS1錶達抑製後PANC1細胞G0/G1期細胞比例明顯升高，而S期和G2/M期細胞比例明顯減小，與對照組比較差異均有統計學意義。結論 SOCS1錶達抑製後，人胰腺癌細胞株PANC1的增殖能力下降，併且對IFN-γ的敏感性增彊。
목적：재인이선암세포중관찰세포인자신호전도억제인자1（SOCS1）침묵대세포증식급간우소-γ（IFN-γ）민감성적영향，탐토SOCS1작위이선암치료파점적가능。방법 Western blot급PCR험증SOCS1간우서렬침묵인이선암세포계PANC1중SOCS1적표체；급여IFN-γ자격후，채용Western blotting방법관찰전록격활인자（STAT）1급린산화STAT（pSTAT）1적변화；채용정량PCR적방법관찰IFN-γ조절인자1（IRF-1）표체수평변화；MTT법검측이선암세포대IFN-γ민감성적변화；세포계수적방법관찰세포적증식속도；채용류식세포술적방법검측세포주기적변화。결과장SOCS1간우서렬전염PANC1세포후，SOCS1 mRNA급단백표체수평균명현하강。침묵SOCS1표체후，pshSOCS1-PANC1조세포적IRF-1 mRNA수평급pSTAT1단백표체수평균현저승고（P<0.05），IFN-γ대PANC1세포적반수억제농도（IC50）현저강저（P<0.01）；전염72 h후PANC1세포수량교대조조현저감소（P<0.05）；SOCS1표체억제후PANC1세포G0/G1기세포비례명현승고，이S기화G2/M기세포비례명현감소，여대조조비교차이균유통계학의의。결론 SOCS1표체억제후，인이선암세포주PANC1적증식능력하강，병차대IFN-γ적민감성증강。
Objective To detect the changes of cell proliferation and IFN-γsusceptibility of human pancreatic can-cer cells after suppressor of cytokine signaling-1 (SOCS1) gene silencing, and to explore the SOCS1 as the target of anti-tu-mor therapy through enhancing the function of IFN-γ. Methods Western blot assay, PCR and real-time PCR were used to verify the down regulation of SOCS1 in human pancreatic cancer cell (PANC1) after transfection;subsequently, PANC1 was stimulated with IFN-γ. Western blot assay was also used to detect the expression of signal transducers and activators of tran-scription (STAT)1 and phosphorylation STAT(pSTAT)1;and the change of IFN-γsusceptibility was detected by MTT assay. Real-time PCR was used to detect the mRNA of interferon regulatory factor-1(IRF-1). Flow cytometry was used to detect the cell cycle. Results The expression levels of SOCS1 mRNA and protein were significantly decreased in small hairpin SCOS1 (shSOCS1) transfected PANC1 cells. After the silence of SOCS1, the expression levels of IRF-1 and pSTAT1 in-creased significantly (P<0.05), and the median inhibitory concentration(IC50)of IFN-γfor PANC1 cells decreased signifi-cantly (P<0.01). The cell count of shSOCS1 cells dropped significantly compared with that of control group after the SOCS1 silencing for 72 hours (P<0.05). The cell cycle arrest was promoted at the G0/G1 phase, but the percentage of cells in S phase and G2/M decreased compared to that of control groups (P<0.05). Conclusion After the inhibition of SOCS1 gene expression, the proliferation ability of human pancreatic cancer cell line PANC1 decreased, and the sensitivity of PANC1 cells to IFN-γwas enhanced.
