探讨JWA基因通过MAPK信号通路对胰腺癌PAN1细胞增殖、凋亡、侵袭和迁移过程的作用.docVIP

目的：探讨JWA基因通过MAPK信号通路对胰腺癌PANC-1细胞增殖、凋亡、侵袭和迁移过程 的作用。 方法：体外培养人胰腺癌PANC-1细胞，针对JWA基因设计干扰RNA(siRNA),并通过脂质体 转染入细胞，用MTT法检测细胞的堉殖能力、用Transwell实验检测细胞的迁移和侵袭能力。 用Western blot免疫印迹分析检测JWA及MAPK信号通路的MEK、Erkl/2、JNK、p38三个通 路中相关蛋白的表达。 结果：通过MTT法我们发现JWA蛋白表达被抑制后，PANC-1细胞的增殖水平有一定升高，面 JWA蛋白表达下调后PANC-1细胞的凋亡水平有一定下降，凋亡相关蛋白BAX的表达显著降低， 凋亡抑制基因Bcl-2的表达有一定的升高。通过Tnmswell实验我们发现JWA siRNA处理后 的PANC-1细胞的穿膜细胞数明显增加，迁移和侵袭能力显著增强。对于JWA与MAPK通路在 胰腺癌细胞系中的关系，我们了解到JWA siRNA组MAPK信号通路中p_MEK和p-Erkl/2蛋白 表达水平有明显的下降，但MEK、Erkl/2、JNK、p38、p-JNK和p-p38的变化不大。 Objective: This thesis is prepared with the objective of discussing the role of JWA gene in the proliferation, apoptosis，invasion and migration of PANC-1 pancreatic cancer cells via MAPK signaling pathways. Methods: Human PANC-1 pancreatic cancer cells were cjiltured in-vitro, and small interfering RNA (siRNA) was designed for JWA gene. Then, siRNA was transfect into the cells via liposome. Cell proliferation was measured via MTT assay，cell migration and invasion was measured via Transwell chambers and protein expression of JWA and MEK, Erkl/2, JNK and p38 of MAPK was measured via Western blot. Results: By utilizing MTT we discovered that when JWA protein expression was inhibited, the proliferation of PANC-1 cells was enhanced to a certain extent. When JWA protein expression was down-regjilated，the apoptosis of PANC-1 cells was decreased to a certain extent，the expression of apoptosis-associated protein (AAP) BAX was substantially decreased and the expression of apoptosis inhibitor gene Bcl-2 was enhanced to a certain extent. Via Transwell chambers，we discovered that the number of penetrating cells of PANC-1 cells was increased dramatically after the PANC-1 cells were treated by JWA siRNA, and the migration and invasion of the cells was substantially enhanced. As to the correlation of JWA and MAPK pathways in pancreatic cancer cells, we came to know that p-MEK and p-Erkl/2 protein expression of MAPK pathway of JWA siRNA control was dramatically down-regjilated，however，the