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LINC01694 regulates the malignant biological behaviors of prostate cancer cells through the miR-128-3p/TERF1 axis
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Abstract:
[Abstract] Objective: To investigate the effects of long non-coding RNA 01694 (LINC01694) regulating the microRNA-128-3p (miR-128-3p)/telomeric repeat binding factor 1 (TERF1) axis on the malignant biological behaviors of prostate cancer (PC) cells. Methods: Cancer tissues and corresponding adjacent tissues from 20 PC patients undergoing surgery at the Department of Urology, Jingzhou Central Hospital, between January 2023 and January 2024 were collected. Human PC cell lines (PC-3, DU145, LNCaP, C4-2) and normal human prostate epithelial RWPE-1 cells were routinely cultured. LNCaP cells were transfected with sh-LINC01694, sh-NC, miR-128-3p inhibitor, inhibitor-NC, miR-128-3pmimics, pcDNA, and pcDNA-LINC01694 using Lipo6000? transfection reagent. Cells were divided into the following groups: Ctrl, sh-NC, sh-LINC01694, sh-LINC01694 + NC inhibitor, sh-LINC01694 + miR-128-3p inhibitor, pcDNA, and pcDNA LINC01694 groups. The mRNA expression of LINC01694, miR-128-3p, and TERF1 in PC tissues and cells, as well as LNCap cells in each group, was detected by qPCR. The protein expression of TERF1, caspase-3, cyclin D1, E-cadherin, and N-cadherin in LNCaP cells of each group was detected by WB method. Clone formation assay, flow cytometry, and Transwell chamber assay were applied to detect proliferation, apoptosis, migration, and invasion of LNCaP cells, respectively. Dual luciferase reporter gene assay, RNA pull-down assay, and RNA-binding protein immunoprecipitation (RIP) assay were applied to verify the targeting binding relationship between LINC01694 and miR-128-3p as well as between TERF1 and miR-128-3p. Nude mouse LNCaP cell xenograft experiment was conducted to assess the effect of LINC01694 knockdown on tumor growth. Results: LINC01694 was highly expressed in PC tissues and cells (all P<0.05). Knockdown of LINC01694 in LNCaP cells promoted the protein expression of miR-128-3p, caspase-3, and E-cadherin, inhibited the protein expression of LINC01694, TERF1, cyclin D1, and N-cadherin, reduced cell proliferation, migration, and invasion, and promoted apoptosis (all P < 0.05). All these effects were partially reversed by the miR 128-3p inhibitor (all P < 0.05). LINC01694 could directly bind to miR-128-3p (P < 0.05), while miR-128-3p could directly bind to TERF1 mRNA (P < 0.05), indicating that LINC01694 regulates the miR-128-3p/TERF1 axis. Knockdown of LINC01694 significantly inhibited the growth of LNCaP cell xenografts in nude mice (P < 0.05). Conclusion: LINC01694 regulates the malignant biological behaviors of LNCaP cells through the miR-128-3p/TERF1 axis.
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