Function Study Of Long Non-coding RNA H19 In Malignant Melanoma

Background Malignant Melanoma?MM?accounts 4%-5%in total malignant tumors and melanoma accounts for approximately 70%death of skin cancer patients.Melanoma is a deadly malignancy and patients with metastatic melanoma had poor prognosis with a 5-year survival rate of about 20%.Though various treatments have been employed for treating melanoma,the most effective ways to treat melanoma were early diagnosis and surgery intervention.Up to date,the exact molecular mechanisms underlying melanoma development,particularly metastasis were largely unknown.Long non-coding RNAs?lnc RNAs?play key roles in the development and progression in some carcinomas which has been reported in recent years.Objective To investigate the role of the long non-coding RNA,H19 in melanoma.Methods In the present study,82 patients with melanoma were included in this study.Melanoma tissues and adjacent normal skin tissues were obtained from patients who underwent surgery at Wuxi No.4 People's Hospital.The expression levels of H19 in clinical samples and melanoma cells were determined by qRT-PCR.The cell growth and cell metastasis were assessed by CCK-8,cell invasion and wound scratching assays.Cell apoptosis and cell cycle were measured by flow cytometry.The effects of H19 on the differential expressions of EMT-related genes were examined by RT²Profiler^TM PCR array.Protein levels were determined by western-blot assay.1205Lu cells stably expressing H19 siRNA2?H19_shRNA2?were constructed,and Xenograft mouse models were performed.Results H19 was highly expressed in melanoma tissues compared to normal adjacent skin tissues,and the tissue expression level of H19 from melanoma patients with metastasis was also significantly higher than that from patients without distant metastasis.In addition,the high expression of H19 in melanoma tissues was associated with shorter overall survival in patients with melanoma.The in vitro functional assays showed that knock-down of H19 inhibited cell growth and metastasis,and also induced cell apoptosis as well as G₀/G₁ arrest.RT² Profiler^TM PCR array and western-blot assay showed that knock-down of H19 differentially regulated the epithelial-mesenchymal transition?EMT?-related gene expressions and reversed EMT in melanoma cell lines.Knock-down of H19 suppressed the in vivo tumor growth in the nude mice.Conclusion In summary,this study may imply that up-regulation of H19 may contribute to the melanoma development,and H19 may be an oncogenic lncRNA for tumor growth and metastasis in melanoma.The findings in the present study may highlight the important role of H19 in melanoma development,and H19 may represent a novel therapeutic target for melanoma in the future.