J Cancer 2019; 10(7):1755-1763. doi:10.7150/jca.28577

Research Paper

Downregulation of DACT-2 by Promoter Methylation and its Clinicopathological Significance in Prostate Cancer

Shibao Li1,2*, Lingyu Yin1,2*, Kai Huang3*, Yao zhao1, Haoliang Zhang1,2, Chenchen Cai1, Yinhai Xu2, Lingyan Huang1, Xiaozhou Wang4, Ting Lan1, Hongchun Li1,2✉, Ping Ma1,2✉

1. School of Medical Technology, Xuzhou Medical University, Xuzhou 221004, China;
2. Department of Laboratory Medicine, Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, China;
3. Department of Urology, Northern Jiangsu People's hospital, Yangzhou 225001 China;
4. The center of functional experiment, Xuzhou Medical University, Xuzhou Jiangsu 221004, China.
*Equal contributors

Abstract

Backgrounds: Dapper homolog (DACT) 2, a member of DACT gene family, is frequently down-regulated in various malignancies and linked to tumor progression. However, the regulatory mechanism of DACT-2 expression and its biological role in human prostate cancer (PCa) remains elusive. Here, we investigated the expression and an epigenetic change of DACT-2 in prostate cancer, and determined if these findings were correlated with clinicopathologic characteristics of PCa.

Methods: The expression profile of DACT-2 of was detected by qRT-PCR, Western blotting, and immunohistochemistry in four prostate cell lines (RWPE-1, LNCaP, PC-3 and DU145), 56 cases of frozen prostate tissues (forty-seven primary prostate carcinomas, nine paired noncancerous and cancerous prostate tissues) and a tissue microarray sets including 100 paraffin-embedded prostate samples (3 normal tissues, 2 cases of adjacent tissues and 95 cases of cancer). Subsequently, the regulatory mechanism of DACT-2 down-regulation was investigated through methylation-specific PCR (MSP) and bisulfite sequencing (BSP). The role of DACT-2 in prostate cancer cell migration and invasion was respectively examined by wound healing and transwell assay. After 5-aza-2'-deoxycytidine treatment of prostate cancer cells, qRT-PCR was used to detect whether the expression of DACT-2 gene mRNA in the cells recovered.

Results: Immunohistochemical results shown that the DACT-2 protein was strongly (3+) expressed in the cytoplasm of all 5 noncancerous tissues and 12.7% (12/95) prostate cancer (PCa) tissues. Whereas 68.4% (65/95) PCa samples and 18.9% (18/95) PCa tissues respectively displayed weakly (1+) expressed and moderately (2+) expressed. In addition, DACT-2 expression was negatively associated with Gleason score in tumor specimens (p=0.029). What's more, down-regulation and promoter methylation of DACT-2 were observed in 68.1% (32/47) frozen PCa tissues and all three prostate cancer cell lines. And, the expression of DACT-2 mRNA was restored by the treatment of demethylated drug 5-aza-2'-deoxycytidine in all prostate cancer lines. Prostate cancer cells invasion and migration were significantly suppressed by ectopic expression of DACT-2 in vitro.

Conclusions: Our study provides evidence that DACT-2 may be a useful biomarker for distinguishing prostate tumor tissues from non-cancerous samples and a potential target for epigenetic silencing in primary prostate Cancer.

Keywords: prostate cancer, DACT-2, methylation, Gleason score, TNM staging

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How to cite this article:
Li S, Yin L, Huang K, zhao Y, Zhang H, Cai C, Xu Y, Huang L, Wang X, Lan T, Li H, Ma P. Downregulation of DACT-2 by Promoter Methylation and its Clinicopathological Significance in Prostate Cancer. J Cancer 2019; 10(7):1755-1763. doi:10.7150/jca.28577. Available from http://www.jcancer.org/v10p1755.htm