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Functional analysis implicating the SNP rs61552325 in ERBB2 as an effector for androgen-insensitive prostate cancer
Xianxiang Xin1,2, Yinmin Gu1, Yang Chen3
1Experimental Centre of Medical Sciences, Guangxi Medical University, Nanning, Guangxi, China.
Background:
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.
Results:
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).
Methods:
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.
Conclusions:
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.
Insights
The ERBB2 rs61552325 variant impacts prostate cancer cell metastasis. The major variant decreases PC3 cell migration, while the minor variant affects DU145 cell invasion.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Prostate cancer is a common malignancy in men.
- Aberrant activation of erb-b2 receptor tyrosine kinase 2 (ERBB2) is critical in metastatic prostate cancer.
- The ERBB2 rs61552325 polymorphism is strongly correlated with prostate cancer risk.
Purpose of the Study:
- To investigate the effect of the ERBB2 rs61552325 polymorphism on androgen-independent prostate cancer cell metastasis.
- To elucidate the functional impact of ERBB2 rs61552325 variants on cancer cell migration and invasion.
Main Methods:
- Site-directed mutagenesis was used to generate major (C allele) and minor (G allele) variants of rs61552325.
- Variants were transfected into DU145 and PC3 prostate cancer cell lines.
- Wound healing and transwell assays were performed to evaluate cell migration and invasion.
Main Results:
- Bioinformatic analysis suggested the Pro1140Ala mutation may decrease ERBB2 protein stability and interactions.
- PC3 cells with the minor variant (G allele) exhibited a 1.28-fold higher migration rate than those with the major variant (C allele).
- DU145 cells with the minor variant showed a significant decrease in invasion compared to controls.
Conclusions:
- The rs61552325 major variant reduces PC3 cell migration.
- The rs61552325 minor variant depresses DU145 cell invasion.
- rs61552325 is a significant genetic factor influencing prostate cancer invasion.