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| LncRNA-TUG1 Affects 22Rv1 Cell Proliferation and Migration by Modulating miR-31-5p/YWHAE Axis |
| ZHAO Jia-Fu*, LI Yong, ZHOU Ming-Shuai, WEN Xiao-Yan, LU Qing-Mei, LIU Bin |
| Guizhou University College of Animal Science/Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang 550025, China |
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Abstract Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activating protein ε (YWHAE) was highly expressed in prostate cancer, and miR-31-5p could target and regulate the expression of YWHAE to affect the proliferation of prostate cancer cells. However, it was not clear which lncRNAs were involved in the targeted regulation of miR-31-5p against YWHAE. In this study, 3 LncRNAs with long non-coding RNA tauronic upregulation gene (LncRNA-TUG1), LINC02604 and PDCD6IP-DT were screened to target and regulate the expression of miR-31-5p based on the results of previous studies and bioanalysis software. The expression of 3 LncRNAs in prostate cancer LNCaP, PC3, 22Rv1 and normal prostate stromal immortem cells (WPMY-1) was detected by RT-qPCR, and the optimal LncRNAs regulating the miR-31-5p/YWHAE axis were screened. The effects of interfering LncRNA-TUG1 and overexpression of miR-31-5p on the expression of LncRNA-TUG1, miR-31-5p and YWHAE in 22Rv1 cells were detected by RT-qPCR. Meanwhile, the effects of co-transfection of sh-TUG1 and miR-31-5p mimics on the proliferation and migration of 22Rv1 cells were detected by CCK-8 and cell scratch assay. Western blot was used to detect the expression of proliferating cell nuclear antigen (PCNA), B cell lymphoma-2 (BCL-2), Bcl-2 associated X protein (BAX) and cysteine proteinase-3 (Caspase-3). The results showed that LncRNA-TUG1, LINC02604 and PDCD6IP-DT were the 3 LncRNAs regulating the miR-31-5p/YWHAE axis, and compared with normal cells, LncRNA-TUG1 was highly expressed in all PCa cell lines, especially in 22Rv1 cells. Taking LncRNA-TUG1 as the research object, after transfecting sh-TUG1 and miR-31-5p mimics into 22Rv1 cells, respectively, qPCR results showed that the expression of miR-31-5p in the miR-31-5p mimics group was significantly up-regulated compared with the NC mimics group (P<0.01). The expression of LncRNA-TUG1 and YWHAE genes was significantly down-regulated (P<0.05). Compared with the sh-NC group, there was no significant change in the expression of miR-31-5p in the sh-TUG1 transfection group, and the expression of LncRNA-TUG1 and YWHAE were significantly down-regulated (P<0.05). CCK-8 and cell scratch experiments showed that co-transfection of sh-TUG1 and miR-31-5p mimics could significantly inhibit cell proliferation and migration. In addition, Western blot results showed that co-transfection of sh-TUG1 and miR-31-5p mimics could significantly inhibit the expression of PCNA and BCL-2/BAX protein, and promoted the expression of caspase-3 protein.These results indicated that LncRNA-TUG1 could regulate the expression of PCNA, BCL-2, BAX and caspase-3 proteins by regulating miR-31-5p/YWHAE, and further affected the proliferation of 22Rv1 cells. This study provides theoretical basis and experimental ideas for the study of targeted drugs for prostate cancer.
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Received: 15 June 2023
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