中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
12期
2496-2499
,共4页
杜涛%张一鸣%郭正辉%陈杰青%周诗丽%赖义明%曹亿%毕良宽%林天歆
杜濤%張一鳴%郭正輝%陳傑青%週詩麗%賴義明%曹億%畢良寬%林天歆
두도%장일명%곽정휘%진걸청%주시려%뢰의명%조억%필량관%림천흠
前列腺特异性膜抗原%p38%前列腺癌%细胞信号通路
前列腺特異性膜抗原%p38%前列腺癌%細胞信號通路
전렬선특이성막항원%p38%전렬선암%세포신호통로
Prostate specific membrane antigen%p38%Prostate cancer%Cell signaling pathway
目的 观察前列腺特异性膜抗原(PSMA)在前列腺癌细胞增殖、迁移和分裂代谢过程中调控相关通路的作用.方法 实验对象包括稳定阻断PSMA表达的细胞株(干扰组)、未阻断PSMA表达的LNCaP细胞株(非干扰组)、不作任何处理的LNCaP细胞株(空白组).利用Western blot及免疫荧光观察3组细胞磷酸化p38(p-p38)的表达量,并使用细胞计数试剂盒(CCK-8)法检测细胞增殖能力,Transwell小室检测细胞迁移能力,流式细胞仪检测细胞周期.结果 Western blot及细胞免疫荧光提示干扰PSMA表达后,p-p38表达水平下降40％(P＜0.05);在SB203582(p38抑制剂)作用下,3组细胞p-p38均处于较低水平,组间比较差异无统计学意义(P＞0.05).CCK-8法、Transwell法、流式细胞仪检测细胞周期提示PSMA干扰后细胞增殖、迁移能力下降,细胞S期百分比降低;给予SB203582后,3组细胞增殖、迁移能力明显下降,细胞S期百分比均处于低水平.结论 PSMA通过上调p38通路对细胞的增殖、细胞周期产生影响,从而对LNCaP细胞起正性调节作用.
目的 觀察前列腺特異性膜抗原(PSMA)在前列腺癌細胞增殖、遷移和分裂代謝過程中調控相關通路的作用.方法 實驗對象包括穩定阻斷PSMA錶達的細胞株(榦擾組)、未阻斷PSMA錶達的LNCaP細胞株(非榦擾組)、不作任何處理的LNCaP細胞株(空白組).利用Western blot及免疫熒光觀察3組細胞燐痠化p38(p-p38)的錶達量,併使用細胞計數試劑盒(CCK-8)法檢測細胞增殖能力,Transwell小室檢測細胞遷移能力,流式細胞儀檢測細胞週期.結果 Western blot及細胞免疫熒光提示榦擾PSMA錶達後,p-p38錶達水平下降40％(P＜0.05);在SB203582(p38抑製劑)作用下,3組細胞p-p38均處于較低水平,組間比較差異無統計學意義(P＞0.05).CCK-8法、Transwell法、流式細胞儀檢測細胞週期提示PSMA榦擾後細胞增殖、遷移能力下降,細胞S期百分比降低;給予SB203582後,3組細胞增殖、遷移能力明顯下降,細胞S期百分比均處于低水平.結論 PSMA通過上調p38通路對細胞的增殖、細胞週期產生影響,從而對LNCaP細胞起正性調節作用.
목적 관찰전렬선특이성막항원(PSMA)재전렬선암세포증식、천이화분렬대사과정중조공상관통로적작용.방법 실험대상포괄은정조단PSMA표체적세포주(간우조)、미조단PSMA표체적LNCaP세포주(비간우조)、불작임하처리적LNCaP세포주(공백조).이용Western blot급면역형광관찰3조세포린산화p38(p-p38)적표체량,병사용세포계수시제합(CCK-8)법검측세포증식능력,Transwell소실검측세포천이능력,류식세포의검측세포주기.결과 Western blot급세포면역형광제시간우PSMA표체후,p-p38표체수평하강40％(P＜0.05);재SB203582(p38억제제)작용하,3조세포p-p38균처우교저수평,조간비교차이무통계학의의(P＞0.05).CCK-8법、Transwell법、류식세포의검측세포주기제시PSMA간우후세포증식、천이능력하강,세포S기백분비강저;급여SB203582후,3조세포증식、천이능력명현하강,세포S기백분비균처우저수평.결론 PSMA통과상조p38통로대세포적증식、세포주기산생영향,종이대LNCaP세포기정성조절작용.
Objective To test the role of prostate specific membrane antigen (PSMA) regulating related pathways in the proliferatation,survival and metabolism of prostate cancer cells.Methods LNCaP cells had been stably transfected with lentivirus-mediated shRNA for PSMA silencing in previous study.The efficacy of PSMA knockdown was testified in LNCaP cell line.By using this PSMA-LNCaP cell line,the expression of PSMA and phospho-p38 (p-p38) detected by using Western blotting was compared among groups.Immunofluorescence was used to confirm the change of p-p38 in cells.Cell viability and migration were measured by cell counting kit-8 reagent and Transwell analysis respectively.Flow cytometry was employed to evaluate the cell survival.P38 pathway inhibitor (SB203582) was used in the culture medium to determine the role of p38 in the prostate cancer cells regulated by PSMA.Results After silencing the expression of PSMA,the expression level of p-p38 was decreased by approximate 40％ as compared with the blank and non-interference groups (P ＜ 0.05).When the cells were incubated with SB203582,the p-p38 expression in three groups was at a low level and no significant difference was found among groups (P ＞0.05).The results of immunofluorescence further proved the relationship between PSMA and p-p38.Decrease of cell viability,migration and survival was observed upon PSMA silencing.SB203580,a specific inhibitor of p38 mitogen-activated protein kinase (MAPK) pathway,also reduced the proliferation,migration and survival of LNCaP cells.Conclusion These data suggest PSMA may stimulate proliferation,migration and survival of prostate cancer cells through p38 MAPK pathway,revealing a novel mechanism for PSMA playing positive roles in LNCaP cells.
