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ADAM17是金属蛋白酶家族(ADAMs)成员之一,研究发现ADAM17可以通过水解细胞表面蛋白的胞外结构域导致肿瘤细胞的增殖和转移.本课题前期研究结果显示,与LNCap细胞相比,ADAM17在DU145细胞中高表达,且与细胞增殖相关.为了研究ADAM17与前列腺癌细胞增殖相关基因p27表达的关系及调控机制,我们采用RNAi技术下调ADAM17的表达,加入PMA(一种ADAM17的激活剂)上调ADAM17的表达,通过细胞计数和CCK-8方法检测细胞增殖,RT-PCR检测p27mRNA的表达,Western印迹检测ADAM17的表达;进一步阻断EGFR和PI3K/Akt信号转导,RT-PCR方法检测p27mRNA的表达,Western印迹检测ADAM17、EGFR、pEGFR、Akt和pAkt的表达.结果显示ADAM17的表达与前列腺癌细胞的增殖呈正相关(P<0.05);p27mRNA的表达与ADAM17的表达呈负相关(P<0.05);分别阻断EGFR和PI3K/Akt信号转导通路,同时使ADAM17表达增加,与对照组(单独PMA处理组)相比,p27mRNA的表达均增加(P<0.05).提示ADAM17调控前列腺癌细胞增殖相关基因p27表达是通过EGFR-PI3K/Akt信号通路实现的.
ADAM17 is a member of the metalloproteinase family (ADAMs), the study found that ADAM17 can cause cell proliferation and metastasis of tumor cells through the hydrolysis of the extracellular domain of cell surface protein.The preliminary results of this topic show that compared with LNCap cells, ADAM17 in DU145 Cells in order to study the relationship between ADAM17 and the expression of p27 in prostate cancer cells and its regulatory mechanism, we down-regulated the expression of ADAM17 using RNAi technology, and added PMA, an activator of ADAM17, up-regulated the expression of ADAM17 The expression of p27 mRNA was detected by RT-PCR and the expression of ADAM17 by Western blotting. The expression of p27 mRNA was further detected by RT-PCR and further by EGFR and PI3K / Akt signaling pathway. Western blot was used to detect the expression of ADAM17, EGFR, pEGFR, Akt and pAkt.The results showed that the expression of ADAM17 was positively correlated with the proliferation of prostate cancer cells (P <0.05), while the expression of p27 mRNA was negatively correlated with the expression of ADAM17 (P <0.05). Respectively, blocked EGFR and PI3K / Akt signal transduction pathways and increased the expression of ADAM17. Compared with the control group (PMA alone), p27 mRNA expression increased (P <0.05). These results suggest that ADAM17 regulates the expression of p27 in prostate cancer cells through the EGFR-PI3K / Akt signaling pathway.