摘要目的 探讨膜联蛋白(Annexin) A2表达下调在前列腺癌侵袭、转移中的作用机制.方法 2010年1月至2012年6月我们采用蛋白质印迹法检测较高转移潜能的前列腺癌细胞C4-2B、较低转移潜能的前列腺癌细胞LNCaP和PC3中Annexin A2表达水平.siRNA技术干扰前列腺癌PC3细胞中Annexin A2表达后,噻唑盐(MTT)法检测细胞增殖情况,流式细胞技术检测细胞凋亡情况,蛋白质印迹法检测基质金属蛋白酶(MMP)-2、MMP-9表达水平的变化,Transwell小室和划痕实验检测前列腺癌PC3细胞体外侵袭能力及迁移能力的变化. 结果 C4-2B细胞中Annexin A2表达水平与内参比较的相对灰度值为0.22,明显低于LNCaP和PC3细胞的0.93和0.95,差异有统计学意义(P＜0.01).将脂质体、Annexin A2的siRNA干扰载体质粒和对照载体质粒分别转染到PC3细胞株,构建3个细胞株:PC3-Lip、PC3-Control Vector、PC3-ANXA2-siRNA.采用siRNA技术干扰Annexin A2表达后,MTT法检测PC3-ANXA2-siRNA细胞生长速度快于PC3、PC3-Lip、PC3-Control Vector细胞,差异有统计学意义(P＜0.05).流式细胞仪检测PC3-ANXA2-siRNA细胞的凋亡率为(6.2±2.6)％,与PC3的(2.3士1.9)％、PC3-Lip的(2.1±2.0)％、PC3-Control Vector的(2.0±1.8)％比较差异无统计学意义(P＞0.05).蛋白质印迹法检测PC3-ANXA2-siRNA细胞的Annexin A2表达水平明显降低,而MMP-2、MMP-9表达水平上调,Transwell小室和划痕实验发现PC3-ANXA2-siRNA细胞体外侵袭及迁移能力增加. 结论 下调Annexin A2的表达能增加前列腺癌细胞的体外侵袭和转移能力,可能的作用机制是上调MMP-2和MMP-9的表达.

abstractsObjective To analyze the impact and its mechanism of down-regulation of AnnexinA2 expression on prostate cancer(PCa) invasion and metastasis.Methods The expression of AnnexinA2 in three prostate cancer cell lines with different metastasis ability,including LNCaP (lower metastasis ability),PC3 (lower metastasis ability),C4-2B (higher metastasis ability) were detected by Western blot.The correlations between the expression of AnnexinA2 and the metastasis ability of prostate cancer cells were also evaluated.The siRNA was used in PC3 cells to down-regulate the expression of AnnexinA2,the cell proliferation assay was performed by MTT method,the cell apoptosis level was detected by flow cytometry,the expression of MMP-2 and MMP-9 were detected by Westrn blot,the in vitro invasiveness of PC3 was detected by transwell cabinet test,and the migration ability of PC3 cells was detected by scratch test,respectively.Results The grayscale value of AnnexinA2 expression in C4-2B cells is 0.22,in contrast with the internal reference,which was obviously lower than those of LNCaP and PC3 cells with lower metastasis potency(relative grey value is 0.93 and 0.95,respectively.P＜0.01).After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,the growth became faster for PC3-ANXA2-siRNA cells than PC3,PC3-Lip and PC3-empty vector cells (P＜0.05).After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,the ratio of apoptosis was detected by flowcytometry in PC3,PC3-Lip,PC3-empty vector and PC3-ANXA2-siRNA cells,and the apoptosis ratio in PC3-ANXA2-siRNA cells was the highest.However,the difference was not significant compare to others (P＞0.05).After RNAi was used in PC3 cells to downregulate the expression of AnnexinA2,the expression of AnnexinA2 in PC3-ANXA2-siRNA cells was decreased while the expression of MMP-2 and MMP-9 were increased.After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,invasiveness of PC3-ANXA2-siRNA cells detected by transwell cabinet test was increased in vitro,and migration of PC3-ANXA2-siRNA cells detected by scratch test was increased in vitro.Conclusions The down-regulation of expression of AnnexinA2 could increase the invasive and metastatic ability of prostate cancer,and this may attribute to the up-regulation of MMP-2 and MMP-9 expression in prostate cancer.