Objective:To observe the expression level of long noncoding RNA RP11-97C16.1 in bladder cancer tissues and its relationship with the survival time of bladder cancer patients, and to explore the role and potential molecular mechanism of RP11-97C16.1 in the proliferation of bladder cancer cells.Methods:The expression difference of RP11-97C16.1 in bladder cancer tissue and adjacent tissue was analyzed by TCGA database, and the relationship between the expression level of RP11-97C16.1 and the survival time of bladder cancer patients was analyzed by GEPIA database. The expression of RP11-97C16.1 in four bladder cancer cell lines (T24, MGH-U3, J82, UM-UC-3) was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The UM-UC-3 cells were divided into RP11-97C16.1 group and control group, and the transfectants were pcDNA-RP11-97C16.1 plasmid and negative control plasmid, respectively. The expression levels of RP11-97C16.1 and miR-3687 were detected by RT-qPCR. The viability and proliferation ability of UM-UC-3 cells were detected by cell counting kit-8 (CCK8) and colony formation assay. The complementary relationship between RP11-97C16.1 and miR-3687 was verified by dual-luciferase reporter gene assay. The expression levels of Cyclin E2, CDK2, CDK4, CDK6 and Cyclin D2 were detected by Western blotting. Measurement data were expressed as mean ± standard deviation ( ± s), independent sample t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:Compared with adjacent tissues, the expression of RP11-97C16.1 in bladder cancer tissues was significantly decreased ( P<0.01). Compared with patients with lower expression of RP11-97C16.1, patients with higher expression of RP11-97C16.1 had longer overall survival time ( P<0.01). Compared with the SV-HUC-1 cell line, the expression of RP11-97C16.1 was significantly decreased in the four bladder cancer cell lines ( P<0.01). In UM-UC-3 cells in which RP11-97C16.1 was upregulated, the expression of miR-3687 was decreased ( P<0.01). Compared with the control group, up-regulation of RP11-97C16.1 could significantly reduce the proliferation ability of UM-UC-3 cells ( P<0.05), and decrease the number of bladder cancer cell colonies ( P<0.01). RP11-97C16.1 could target and bind miR-3687 ( P<0.01). Compared with the control group, overexpression of RP11-97C16.1 could significantly decreased the expression of Cyclin E2, CDK2, CDK4, CDK6, and Cyclin D2 proteins. Conclusions:The expression of RP11-97C16.1 is low in bladder cancer tissue, and patients with higher expression of RP11-97C16.1 have a longer survival time. Up-regulation of RP11-97C16.1 can down-regulate the expression of miR-3687, thereby inhibiting the proliferation of bladder cancer cells UM-UC-3.

Objective:To explore the expression of microRNA (miRNA)-6516-5p in renal cancer cell lines and the molecular mechanisms regulating the proliferation and migration of renal cancer cells.Methods:quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-6516-5p in renal cancer cell lines and normal proximal renal tubular epithelial cell lines. The liposome method was used to transiently transfect miR-6516-5p mimic and nonsense sequence (NC) into renal cancer cells with the lowest expression of miR-6516-5p, namely miR-6516-5p group and NC group. qRT-PCR was used to detect the expression of miR-6516-5p in transfected cells. CCK-8 and Transwell migration experiment were used to detect the proliferation and migration of transfected cells. Bioinformatics software and dual luciferase gene report experiment were used to predict and verify the regulation of miR-6516-5p on target gene, respectively. qRT-PCR and Western blotting were used to detect the expression of target gene in transfected cells. Measurement data were expressed as mean±standard deviation ( ± s), t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:The expression of miR-6516-5p in renal cancer cell lines was significantly lower than that of normal proximal tubular epithelial cells ( P<0.01), and the expression of miR-6516-5p in 786-O cells was the lowest ( F=27.69, P<0.01). The expression of miR-6516-5p in 786-O cells in NC group and miR-6516-5p group was 1.01±0.08 and 9.91±1.16, respectively. Compared with the NC group, the expression of miR-6516-5p in 786-O cells in the miR-6516-5p group was significantly increased ( t=7.63, P<0.01). Up-regulation of miR-6516-5p can significantly inhibit the proliferation of 786-O cells ( P<0.05). The migration numbers of NC group and miR-6516-5p group were 85.65±8.77 and 28.05±6.20, respectively. Overexpression of miR-6516-5p could inhibit the migration of 786-O cells ( t=5.36, P< 0.01). The target gene of miR-6516-5p may be ornithine decarboxylase 1 ( ODC1), miR-6516-5p can significantly inhibit the luciferase activity of wild-type ODC1-3′UTR ( t=9.83, P<0.01). Up-regulation of miR-6516-5p can reduce the expression of ODC1 mRNA and protein in 786-O cells ( P<0.01). Conclusion:The expression of miR-6516-5p is reduced in renal cancer cell lines, miR-6516-5p inhibits the proliferation and migration of renal cancer 786-O cells by targeting ODC1, miR-6516-5p may become a potential molecular target of renal cancer.

Objective:To investigate the mechanism of physcion affecting the cell cycle and proliferation of prostate cancer DU145 cell line by regulating the expression of miR-380-3p.Methods:Prostate cancer DU145 cells were treated with 50 μg/mL physcion as physcion group, and normal cultured DU145 cells without any treatment were used as control group. Flow cytometry was used to detect DU145 cell cycle changes. MTT proliferation test was used to detect the proliferation of DU145 cells. quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-380-3p in DU145 cells. The bioinformatics software RNAhybrid was used to predict the target genes of miR-380-3p. qRT-PCR and Western blotting methods were used to detect the expression of miR-380-3p target gene. Measurement data were expressed as mean ± standard deviation ( ± s), t-test was used for comparison between two groups. Results:Compared with the control group, DU145 cells in the physcion group were blocked in the G 0/G 1 phase ( P<0.01), and the proliferation ability of DU145 cells was significantly inhibited ( P<0.05). The expression of miR-380-3p in DU145 cells in the control group and physcion group was 8.36 ± 1.42 and 1.08 ± 0.39, respectively. Physcion could promote the expression of miR-380-3p ( t=4.96, P<0.01). The functional target gene of miR-380-3p may be UHRF1. The relative expression levels of UHRF1 mRNA in DU145 cells in the physcion group and control group were 0.23±0.06 and 1.04±0.15, respectively. Compared with the control group, the expression of UHRF1 gene in DU145 cells in the physcion group was decreased ( t=4.55, P<0.01). Conclusion:Physcion can inhibit the proliferation of prostate cancer DU145 cells and induce G 0/G 1 block in DU145 cells, which may be closely related to the regulation of miR-380-3p.

Objective:To explore the mechanism by which microRNA (miRNA) -1303 inhibits the proliferation and migration of renal cell carcinoma 786-O cells through targeted regulation of lysophosphatidic acid receptor 3 (LPAR3) expression.Methods:quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression of miR-1303 in renal cancer cell lines (A498, ACHN, 786-O, OS-RC-2) and normal renal tubular epithelial cells HK-2. The miR-1303 mimic and the negative control sequence were transfected into the renal cancer cells with the lowest expression of miR-1303, respectively, as the miR-1303 group and the negative control group. qRT-PCR detected the relative expression of miR-1303 in the two groups of cells. MTT method and Transwell migration experiment were used to detect cell proliferation and migration ability. RegRNA 2.0 predicted the target genes of miR-1303. The dual luciferase reporter gene detected the binding of miR-1303 to the target gene. qRT-PCR and Western blotting detected the relative expression of LPAR3. Measurement data were expressed as mean±standard deviation ( ± s), t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:The expressions of miR-1303 in renal cancer cell lines A498, ACHN, 786-O, OS-RC-2 and normal renal tubular epithelial cells HK-2 were 0.51±0.04, 0.79±0.02, 0.21±0.04, 0.55±0.07 and 1.00±0.05, the expression of miR-1303 in renal cancer cell lines was lower than that in HK-2 ( P<0.05), and the relative expression in 786-O cells was the lowest ( F=29.50, P<0.01). Compared with the control group, the expression of miR-1303 in the experimental group was significantly increased [(1.00±0.01) vs (7.98±0.88), t=7.95, P<0.01]. The cell absorbance value of the experimental group was significantly lower than that of the control group ( P<0.05). The number of cell migration in the experimental group was significantly lower than that in the control group ( P<0.05). miR-1303 can bind to LPAR3 mRNA in a complementary pair ( P<0.01). Compared with the control group, the expression of LPAR3 mRNA in the 786-O cells of the experimental group was significantly reduced [(1.00±0.01) vs (0.23±0.03), t=23.56, P<0.01]. Conclusion:miR-1303 may inhibit the proliferation and migration ability of renal cancer 786-O cells by down-regulating the expression of LPAR3.

Objective:To explore the expression level of miR-769-3p in bladder cancer tissues, and observe the effect of silencing miR-769-3p on the migration ability and cell cycle of J82 cells by down-regulating the expression level of miR-769-3p in bladder cancer J82 cells.Methods:The OncomiR database was used to analyze the expression differences of miR-769-3p in bladder cancer tissues and adjacent tissues. J82 cells were transfected with Lipofectamine 2000 transfection reagent and divided into si-miR-769-3p group (transfected with miR-769-3p small molecule interference fragments) and control group (transfected with meaningless sequences). quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-769-3p after transfection. The cell scratch test and flow cytometry were used to compare the migration ability and cell cycle differences between the two groups of J82 cells. The bioinformatics software MicroRNAdb was used to predict the target gene of miR-769-3p. The dual-luciferase reporter gene assay was used to verify the complementary binding of miR-769-3p to the target gene. qRT-PCR and Western blotting were used to detect the expression levels of miR-769-3p target gene. Measurement data were expressed as mean ± standard deviation ( ± s), and t-test was used for comparison between two groups. Results:The expression of miR-769-3p was significantly increased in bladder cancer tissues compared with adjacent tissues, the difference was statistically significant ( P<0.01). The relative expression of miR-769-3p in the si-miR-769-3p group (1.02 ± 0.16) was significantly lower than that of the control group (4.50 ± 0.60), the difference was statistically significant ( P<0.01). The cell migration rate of the si-miR-769-3p group [(26.67±3.98)%] was significantly lower than that of the control group [(61.86±4.70)%], the difference was statistically significant ( P<0.01). The proportion of cells in the G 0-G 1 phase in the si-miR-769-3p group [(57.66±5.74)%] was significantly higher than that in the control group [(31.26±3.24)%], the difference was statistically significant ( P<0.01). Dual-luciferase reporter gene assay confirmed that endothelin 3 ( EDN3) was the target gene of miR-769-3p. The relative expression of EDN3 mRNA in J82 cells in control group and si-miR-769-3p group was 1.99 ± 0.66 and 6.98 ± 0.76, compared with the control group, the EDN3 mRNA relative expression level of the si-miR-769-3p group was significantly higher than that of the control group, the difference was statistically significant ( P<0.01). Conclusion:Low expression of miR-769-3p can inhibit the migration of bladder cancer J82 cells and block the J82 cell cycle by promoting the expression of EDN3 gene.

Objective:To investigate the effects of astragalin on the cell proliferation and cell cycle of prostate cancer cell line C4-2B through up-regulating the expression of miRNA-513 (miR-513).Methods:Prostate cancer cell line C4-2B cells were taken and treated with 125 μg/L of astragalin for 48 h (astragalin group), and untreated C4-2B cells were set as the control group. The methyl thiazolyl tetrazolium (MTT) method was used to detect the proliferation ability of C4-2B cells in the two groups, and cell cycle was detected by using flow cytometry. The miRNAMap prediction software was used to predict that the targeted gene of miR-513 was the forkhead box protein R2 (FOXR2), and the dual luciferase gene reporter assay was used to verify it. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-513 and FOXR2 mRNA in the two groups of cells. Western blotting was used to detect the expressions of FOXR2, cyclin-dependent kinase 7 (CDK7), β-actin and cyclin H in the two groups of C4-2B cells.Results:Compared with the control group, the proliferation activity of C4-2B cells in the astragalin group was decreased from day 2 to day 5 (all P < 0.05). The proportions of S-phase cells in the control group and the astragalin group were (48.1±3.2)% and (36.0±2.1)%, respectively. The proportion of S-phase cells in the astragalin group was decreased ( t = 3.12, P = 0.021); the proportions of G 2-phase cells were (24.9±3.3)% and (11.8±2.4)%, respectively. The proportion of G 2-phase cells in the astragalin group was decreased ( t = 3.18, P = 0.019). The relative expression levels of miR-513 in C4-2B cells of the control group and the astragalin group were 1.01±0.22 and 6.55±0.61, respectively. The relative expression levels of miR-513 in C4-2B cells in the astragalin group was increased ( t = 7.70, P < 0.01). The dual luciferase reporter gene assay verified that FOXR2 was the targeted gene of miR-513. The relative expression level of FOXR2 mRNA in C4-2B cells of the control group and the astragalin group was 1.04±0.14 and 0.19±0.06, respectively, and the difference was statistically significant ( t = 5.53, P = 0.002), suggesting that after astragalin promoted the expression of miR-513, the FOXR2 mRNA expression was decreased. The relative expression levels of FOXR2, CDK7 and cyclin H protein in C4-2B cells in the astragalin group were all decreased compared with those in the control group. Conclusions:Astragalin inhibits the proliferation of prostate cancer C4-2B cells and induces cell cycle arrest by up-regulating the expression of miR-513.

Objective:To explore the effect of long non-coding RNA (lncRNA) AC068768.1 on the cycle and proliferation of renal cancer cells and its molecular mechanism.Methods:Real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression of AC068768.1 in renal cancer cell lines. The OS-RC-2 cells with the lowest expression of AC068768.1 were used as the transfection objects, OS-RC-2 transfected with the negative control plasmid was set as the control group, and the cells transfected with the AC068768.1 plasmid were set as the AC068768.1 group. qPCR was used to detect the expression of AC068768.1 in transfected OS-RC-2 cells. The effects of AC068768.1 on the cell cycle and proliferation of OS-RC-2 were detected by flow cytometry and tetramethylazazole blue colorimetric (MTT) proliferation experiments. Using bioinformatics methods to predict the microRNA (miRNA) that AC068768.1 may bind. qPCR was used to detect the expression of miRNA and downstream gene mRNA, and Western blot was used to detect the expression of downstream gene protein.The measurement data were expressed as mean±standard deviation ( Mean± SD), the comparison between the two groups adopts the t-test, and the comparison among multiple groups adopts the One-way analysis of variance. Results:Compared with normal renal tubular epithelial cells, the expression of AC068768.1 in renal cancer cell lines was significantly reduced, the difference was statistically significant ( P<0.01). The expression of AC068768.1 in OS-RC-2 cells in the AC068768.1 group was significantly higher than that in the control group, the difference was statistically significant ( P<0.01). Up-regulating the expression of AC068768.1 can inhibit the cycle ( P<0.05) and proliferating ability ( P<0.05) of renal cancer cells. miR-21-5p may be the functional target gene of AC068768.1. Up-regulation of AC068768.1 can significantly inhibit the expression of miR-21-5p ( P<0.01) and promote the expression of tissue inhibitor of metalloproteinase 3 (TIMP3) ( P<0.01). Conclusion:AC068768.1 promotes the expression of TIMP3 gene by regulating the expression of miR-21-5p, thereby inhibiting the cell cycle and proliferation of renal cancer OS-RC-2 cells.

Objective:To observe the expression of long non-coding RNA (lncRNA) PEBP1P2 in renal cell carcinoma (RCC) tissues and its effect on the proliferation and migration of RCC cells.Methods:The expression of PEBP1P2 in 51 RCC tissues and RCC cell lines was detected by real-time quantitative polymerase chain reaction (qPCR). The A498 cells with the lowest expression of PEBP1P2 were transfected, and the cells transfected with PEBP1P2 plasmid were used as the PEBP1P2 group, and the cells transfected with the negative control plasmid were used as the NC group. qPCR was used to detect the expression of PEBP1P2 in the two groups of cells. MTT assay and Transwell migration assay were used to detect the proliferation and migration ability of RCC cells. qPCR and Western blotting were used to detect the expression of caspase recruitment domain family member 10 ( CARD10) gene and NF-κB pathway protein, respectively. Measurement data were expressed as mean±standard deviation ( Mean± SD), and LSD- t test was used for comparison between groups. Results:The expression of PEBP1P2 in RCC tissues was lower than that in adjacent tissues ( t=4.89, P<0.01). The expression of PEBP1P2 in RCC cells was lower than that in normal renal tubular epithelial cells ( P<0.01). The expression of PEBP1P2 in A498 cells of the PEBP1P2 group and NC group was (11.01±1.26) and (1.06±0.19), respectively, and the PEBP1P2 group was significantly higher than that in the NC group ( t=7.81, P<0.01). Overexpression of PEBP1P2 significantly inhibited the proliferation of RCC cells ( P<0.05) and migration ability ( t=3.65, P<0.05). Overexpression of PEBP1P2 significantly suppressed the expression of CARD10 gene in RCC A498 cells ( t=6.83, P<0.01) and inhibited the transduction of NF-κB signaling pathway proteins. Conclusions:PEBP1P2 expression was significantly decreased in RCC tissues. Overexpression of PEBP1P2 significantly inhibited the proliferation and migration of RCC A498 cells. Its molecular mechanism is that PEBP1P2 down-regulates CARD10 gene expression and inhibits NF-κB signaling pathway.

Objective: To investigate the effect of long-chain non-coding RNA AL117378.1 on the proliferation and invasion of bladder cancer cells. Methods: From August 2016 to December 2017, the tumor tissues and adjacent tissues of 14 patients with bladder cancer treated by Huangshi Central Hospital of Edong Healthcare Group were selected. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of AL117378.1 in 14 pairs of bladder cancer tissues and corresponding adjacent tissues, bladder cancer cell lines (5637, J82, T24, BIU-87) and corresponding human bladder normal epithelial cells (SV-HUC-1). The negative control plasmid (control group) and the plasmid carrying the AL117378.1 (experimental group) were transfected into the bladder cancer cell line with the lowest expression level. qRT-PCR was used to detect the expression levels of AL117378.1 and non-receptor protein tyrosine phosphatase 9 (PTPN9) mRNA in the transfected cells. The expression levels of PTPN9, E-cadherin, α-SMA, CDK4 and Cyclin A2 protein were detected by Western blotting. MTT assay and Transwell invasion assay were used to detect the cell proliferative capacity and invasive ability of the two groups. Measurement data were expressed as mean± standard deviation(Mean±SD), and comparison between groups used independent sample t test. Results: The expression of AL117378.1 in bladder cancer tissues was lower than that in adjacent tissues (P<0.01). The expression of AL117378.1 in bladder cancer cell lines was lower than that in human bladder normal epithelial cells (P<0.01), and the expression level of AL117378.1 was the lowest in J82 cells (P<0.01). The expression levels of AL117378.1 in the control and experimental groups were 1.02 ± 0.11 and 7.96 ± 1.06, respectively, and the difference was statistically significant (t=6.51, P<0.01). The expression of PTPN9 mRNA was 1.01 ± 0.08 and 4.99 ± 0.50, respectively. the difference was statistically significant (t=7.84, P<0.01). Western blotting results showed that the expression of PTPN9 and E-cadherin increased, and the expression of α-SMA, CDK4 and Cyclin A2 decreased. MTT experiments showed that the proliferation of cells transfected with AL117378.1 was significantly decreased (P<0.05). Transwell invasion experiments showed that the number of invasive cells in the control group and the experimental group were 97.96±13.71 and 38.02±7.51, respectively, and the difference was statistically significant (t=3.84, P<0.01). Conclusions The expression of AL117378.1 was significantly decreased in bladder cancer tissues and cell lines (5637, J82, T24, BIU-87). AL117378.1 can significantly inhibit the proliferation and invasion of bladder cancer cells by positively regulating the expression of PTPN9 gene.

Objective:To investigate the expression of long non-coding RNA (lncRNA) COX10-AS1 in renal cell carcinoma tissues and cell lines and its effect on proliferation and migration of renal cancer cells.Methods:Fluorescence real-time quantitative PCR (qRT-PCR) was used to detect the expression of COX10-AS1 in surgical specimens that have been diagnosed as renal cancer tissues and adjacent tissues by pathology, renal cancer cell lines (786-O, CaKi-1, A498, ACHN) and normal renal tubular epithelium cell line (HK-2). The ACHN cells with the lowest expression were divided into a control group (transfected with a negative control plasmid carrying nonsense sequences) and an experimental group (transfected with a plasmid carrying COX10-AS1 sequences). The expression level of COX10-AS1 was detected by qRT-PCR in two groups of cells. The proliferation and migration ability of ACHN cells were detected by MTS assay and cell scratch assay. The expression of MFN2 mRNA was detected by qRT-PCR. The expressions of MFN2 and Ras-NF-κB signaling pathway proteins were detected by Western blotting. The measurement data were expressed as mean±standard deviation ( Mean± SD), the comparison between the two groups used the t-test, and the comparison among multiple groups adopts the one-way analysis of variance. Results:The expression of COX10-AS1 in renal cell carcinoma was significantly lower than that in adjacent tissues ( P<0.01), The expression of COX10-AS1 in renal cell carcinoma cells was significantly lower than that in renal tubular epithelial cells ( P<0.05), the expression of COX10-AS1 was the lowest in ACHN cells( P<0.01), the above differences were statistically significant compared with the control group, the expression of COX10-AS1 in ACHN cells of experimental group was significantly increased ( P<0.01), the above differences were statistically significant compared with the control cells, the proliferation of ACHN cells in the experimental group was significantly decreased ( P<0.05), and the cell migration ability was significantly decreased ( P<0.01). Compared with the control cells, the expression of MFN2 mRNA in ACHN cells of experimental group was significantly increased ( P<0.01). The expression levels of MFN2 were significantly up-regulated ( P<0.01), and Ras-NF-κB signaling pathway proteins were significantly down-regulated ( P<0.05), the above differences were statistically significant. Conclusions:The expression of COX10-AS1 is decreased in renal cell carcinoma tissues and cell lines. COX10-AS1 may inhibit the proliferation and migration of ACHN cells by promoting the expression of MFN2 gene.