Xin Xianxiang, Gu Yinmin, Chen Yang, Huang Yuanjie, Mo Zengnan, Hu Yanling
Experimental Centre of Medical Sciences, Guangxi Medical University, Nanning, Guangxi, China.
Commission for Discipline Inspection, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, China.
Oncotarget. 2017 May 16;8(20):33745-33755. doi: 10.18632/oncotarget.16807.
As one of the most common cancers in men, the pathogenesis of prostate cancer has been widely researched. Aberrant activation of the erb-b2 receptor tyrosine kinase 2 (ERBB2) has been found to play a critical role in metastatic prostate cancer. In our previous study, we demonstrated that rs61552325 (Pro1140Ala) located in ERBB2 is strongly correlated to prostate cancer. Therefore, we initially studied the effect of rs61552325 on androgen-independent prostate cancer cell metastasis.
Bioinformatic results demonstrated that the mutant Pro1140Ala likely decrease the stability of the ERBB2 protein and its interactions. The mean migration rate after 6 h for PC3 minor variant cells which carried the G allele was 1.28-fold higher than major variant PC3 cells that carried the C allele (P = 0.016). The mean invasion rate of DU145 putative minor variant cells was 0.40 reducer than negative control cells (P = 5.9E-04).
rs61552325 major variant (C allele) and minor variant (G allele) were produced by site directed mutagenesis and transfected into DU145 and PC3 cells. A wound healing assay was performed to compare migration abilities between alleles. After knocking down endogenous ERBB2 and then expressing the rs61552325 minor variant, invasion abilities were evaluated with a transwell assay using DU145 and PC3 cells.
Our data showed that the rs61552325 major variant decreases PC3 cell migration and its minor variant depresses DU145 cell invasion, suggesting that rs61552325 is likely an important change during prostate cancer invasion.
前列腺癌作为男性最常见的癌症之一，其发病机制已得到广泛研究。已发现erb-b2受体酪氨酸激酶2（ERBB2）的异常激活在转移性前列腺癌中起关键作用。在我们之前的研究中，我们证明位于ERBB2中的rs61552325（Pro1140Ala）与前列腺癌密切相关。因此，我们最初研究了rs61552325对雄激素非依赖性前列腺癌细胞转移的影响。
生物信息学结果表明，突变体Pro1140Ala可能会降低ERBB2蛋白的稳定性及其相互作用。携带G等位基因的PC3次要变体细胞在6小时后的平均迁移率比携带C等位基因的主要变体PC3细胞高1.28倍（P = 0.016）。DU145推定次要变体细胞的平均侵袭率比阴性对照细胞低0.40（P = 5.9E-04）。
通过定点诱变产生rs61552325主要变体（C等位基因）和次要变体（G等位基因），并将其转染到DU145和PC3细胞中。进行伤口愈合试验以比较等位基因之间的迁移能力。在敲低内源性ERBB2然后表达rs61552325次要变体后，使用DU145和PC3细胞通过Transwell试验评估侵袭能力。
我们的数据表明，rs61552325主要变体降低了PC3细胞的迁移能力，其次要变体抑制了DU145细胞的侵袭能力，这表明rs61552325可能是前列腺癌侵袭过程中的一个重要变化。
