Chinese Journal Of Cell And Stem Cell >Archive >2020 >Volume 10 Number 3 June 2020 >Original Researches >contents

Journal content

Effects of lncRNA RPL34-AS1 on the proliferation and apoptosis of ovarian cancer cells by targeted regulation of miR-575 expression

Zhang Jing, Dong Jie, Wang Qiuju, et al


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【Abstract】 Objective To investigate the effect of RPL34-AS1 on the proliferation and apoptosis of ovarian cancer cells and whether its mechanism is related to miR-575. Method SKOV3 cells in logarithmic growth phase were synchronized with serum-free mediumfor 12 h and pcDNA, pcDNA-RPL34-AS1, si-NC, si-NC, si-RPL34-AS1, si-RPL34-AS1, anti-miR-NC, anti-anti-miR- NC, anti-miR-575 were transfected into SKOV3 cells, respectively labeled as pcDNA group, pcDNA- RPL34-AS1 group, si-NC group, si-NC group, si-RPL34-AS1 group, si-RPL34- AS1 group, anti-miR-NC group, anti-miR-NC group, anti-miR-575 group; PcDNA-RPL34- AS1 was co- transfected into SKOV3 cells with miR-NC and miR-575, respectively, labeled as pcDNA- RPL34- AS1+miR-NC group, pcDNA-RPL34-AS1+miR-575 group. Real-time fluorescent quantitative PCR (RT-qPCR)was used to detect the expression level of RPL34-AS1 and miR-575 in clinical tissue specimens and cells of each group after transfection; dual luciferase reporter assay was used to detect the targeting relationship between RPL34-AS1 and miR-575; tetramethylazozolium salt colorimetric method (MTT) was used to detect cell viability; flow cytometry was used to detect apoptosis; Western Blot was used to detect expression of Cyclin D1, Cyclin-dependent kinase inhibitor 1A (p21), B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2 related X protein (Bax) proteins.The comparison between the two groups was performed using the independent sample t test, and the comparison between multiple groups was performed using the one-way analysis of variance; LSD-t test was used for pairwise comparison between groups. Results Compared with adjacent tissues, the expression level of RPL34-AS1 was decreased in ovarian cancer tissues, the expression level ofmiR-575 was increased (1.01±0.07 vs 3.12±0.28) (P < 0.05). After transfection with si-RPL34-AS1, cell viability was increased (48 h: 0.68±0.06 vs 0.55±0.05; 72 h: 0.99±0.08 vs 0.71±0.06), and the proportion of G1 phase cells was decreased (13.42±1.38 vs 32.15±2.11), the proportion of S-phase cells was increased (53.75±5.22 vs 34.69±3.41), the apoptosis rate was decreased (4.31±0.42 vs 9.25±0.91), and the expression level of CyclinD1 and Bcl-2 were increased (0.92±0.08 vs 0.71±0.07; 0.86±0.07 vs 0.61±0.06), the expression level of p21 and Bax were decreased (0.13±0.02 vs 0.29±0.03; 0.19±0.02 vs 0.31±0.03) (all P < 0.05). RPL34-AS1 targets and regulates miR-575. Overexpression of RPL34-AS1 or inhibition of miR-575 can inhibit cell activity, block the cell cycle and promote apoptosis. MiR-575 overexpression reversed RPL34-AS1 overexpression effect on ovarian cancer SKOV3 cell proliferation inhibition as well as its apoptosis promotion. Conclusion Overexpression of RPL34-AS1 can inhibit the proliferation of ovarian cancer SKOV3 cells and promote cell apoptosis, which may be related to the down-regulation of miR- 575.
 

【Key words】 RPL34-AS1; MiR-575; Ovarian cancer; Proliferation; Apoptosis