Pttg1和MMP13调控前列腺癌细胞侵袭力的研究

Regulative effect of pituitary tumor-transforming gene 1 and matrix metalloproteinase 13 on invasion of prostate cancer cells

目的:了解垂体肿瘤转化基因1(Pttg1)和基质金属蛋白酶13(MMP13)与前列腺癌(PCa)的关系以及二者在PCa转移中所起的作用。方法:通过RT-qPCR法了解PCa组织中Pttg1和MMP13表达水平及二者的相关性。分别使用过表达Pttg1的质粒或携带shRNA的质粒(shPttg1)转染人PCa细胞系PC3来上调或下调Pttg1水平,测定相应的MMP13表达水平的变化(用RT-qPCR、ELISA法来测定);反之测定MMP13表达水平的变化对Pttg1的影响。用划痕愈合实验或Transwell细胞迁移实验测定肿瘤细胞侵袭的变化。用特异性通路抑制剂抑制ERK/MAPK、JNK、PI3K信号通路,并用Western Blot法测定这些通路抑制剂在Pttg1过表达的PC3细胞中对MMP13水平的影响效果。结果:本研究发现,在病理学组织中,PCa组织中Pttg1和MMP13的表达水平显著高于邻近正常前列腺组织,并且Pttg1和MMP13的表达水平具有相关性。体外实验中,Pttg1激活MMP13,而后者决定PCa细胞的侵袭力。但Pttg1表达水平不受MMP13的显著影响。另外,PCa细胞中Pttg1激活的MMP13可被PI3k/Akt信号通路显著抑制,而不是ERK/MAPK或JNK信号通路。结论:本研究显示,PCa组织中Pttg1和MMP13的表达水平显著增高,二者有相关性;Pttg1可通过MMP13增加PCa细胞的侵袭能力,提示Pttg1/MMP13轴有望成为PCa治疗的靶点。

Objective：To investigate the relationship between pituitary tumor-transforming gene 1（Pttg1）,matrix metalloproteinase 13（MMP13）and prostate cancer（PCa）as well as their roles in the metastases of PCa.Method：Pttg1and MMP13 levels and the correlation were detected in PCa specimens by RT-qPCR.We used either a Pttg1-overexpressing plasmid or a plasmid carrying short hairpin small interfering RNA（shRNA）for Pttg1（shPttg1）to transfect a human PCa cell line,PC3,so as to increase or decrease Pttg1 levels,respectively.Also,corresponding MMP13 levels were detected by RT-qPCR and ELISA.In reverse,the effects of MMP13 modification on Pttg1 were detected.Cell invasion was tested in either a scratch wound healing assay or a transwell cell migration assay.We used specific inhibitors to inhibit ERK/MAPK,JNK and PI3 Ksignaling pathway and checked the effects of application of these specific pathway inhibitors on the MMP13 levels in Pttg1 overexpressed PC3cells by Western Blot.Result：We found significantly higher levels of Pttg1 and MMP13in the resected PCa specimens,compared with the adjacent normal prostate tissues from the same patient.Pttg1 and MMP13levels strongly correlated with each other.In vitro,Pttg1 activated MMP13,which determined PCa cell invasion.However,Pttg1 levels were not significantly affected by MMP13.Furthermore,the Pttg1-activated MMP13 in PCa cells was significantly suppressed by inhibition of PI3k/Akt,but not ERK/MAPK or JNK pathways.Conclusion：Our data suggest that Pttg1 and MMP13levels significantly increase and correlate with each other in PCa specimens.Pttg1 may increase PCa cell metastasis by MMP13,so Pttg1/MMP13 axis can be regarded as a promising therapeutic target for PCa treatment.