Objective:By observing the effects of Calycosin on the proliferation and migration of human renal cancer 769-P cell, to explore the possible molecular mechanism of Calycosin against renal cancer.Methods:769-P cell were cultured with different concentrations of Calycosin [0, 12.5, 25, 50, 100, 200 μmol/L, dissolved in Dimethyl sulfoxide (DMSO)], and the effects of different concentrations of Calycosin on the viability of 769-P cell was detected by CCK8 method. The 769-P cell treated with 200 μmol/L Calycosin were used as the Calycosin group, and the 769-P cell treated with DMSO were used as the control group. The cell colony formation assay and cell scratch assay were used to detect the effects of Calycosin on the proliferation and migration of 769-P cell, respectively. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the effect of Calycosin on the expression of miRNA-1246 and chemokine receptor-4 (CXCR4) in 769-P cell. Western blotting method was used to detect the effects of Calycosin on the expression of CXCR4 and extracellular signal-regulated kinase (ERK) pathway proteins in 769-P cell. Measurement data were expressed as mean ± standard deviation ( ± s), and one-way ANOVA was used for comparison between multiple groups, while t-test was used for comparison between two groups. Results:After cultured with 0, 12.5, 25, 50, 100, and 200 μmol/L of Calycosin, the absorbance values of renal cancer 769-P cell were 0.99 ± 0.06, 0.74 ± 0.07, 0.60 ± 0.03, 0.55 ± 0.05, 0.40 ± 0.06, 0.21 ± 0.04, respectively; compared with 0 μmol/L, the Calycosin could reduce the survival rate of 769-P cell ( P<0.05). The number of clones of 769-P cell in the control group and the Calycosin group was 109.80 ± 13.19 and 60.66 ± 11.22, respectively, and the number of clones of the 769-P cell in the Calycosin group was decreased, the difference was statistically significant ( t=5.67, P<0.01). The relative migration rates of 769-P cell in the control group and the Calycosin group were (43.13 ± 3.82)% and (14.27 ± 3.25)%, respectively, after the 769-P cell were treated with Calycosin, the cell migration ability was weakened ( t=5.71, P<0.05). The relative expression levels of miRNA-1246 in 769-P cell of the control group and the Calycosin group was 1.03 ± 0.12 and 6.99 ± 1.84, respectively, and the relative expression levels of CXCR4 mRNA was 7.17 ± 2.96 and 0.98 ± 0.06, respectively, showed that Calycosin can up-regulate the expression of miRNA-1246 in 769-P cell ( t=3.24, P<0.01), and down-regulate the expression of CXCR4 mRNA ( t=4.18, P<0.01). Compared with the control group, the Calycosin could down-regulate the expression of CXCR4 protein and ERK pathway protein in 769-P cell. Conclusion:Calycosin can inhibit the proliferation and migration of renal cancer 769-P cell, and its mechanism may be related to up-regulating the expression of miRNA-1246 and blocking the CXCR4/ERK pathway.

During orthodontic treatment,irregular and varying sized nodular bony protrusions may sometimes appear on the labial side of the patient's anterior alveolar bone,which is closely related to the differential bone remodeling on the inner and outer surfaces of the alveolar bone.Labial protuberances not only affect the aesthetic results of orthodontic treatment,but also pose potential risks to periodontal health.Currently,it is believed that the influencing factors of the formation of the labial protuberances may be related to the patient's gender and age,tooth movement speed,and extent of anterior teeth retraction.Labial protuberances typically resolve spontaneously,however,if persistent,alveoloplasty may be necessary for treatment.This review provides a summary on the occurrence,influencing factors of formation,potential biological mechanisms,and corresponding treatment methods of labial protuberances during orthodontic treatment.

Objective:To explore the effect of miR-1249-5p on the proliferation, metastasis and cell cycle of PC-3 cell in prostate cancer.Methods:The relationship between the expression level of miR-1249-5p and the overall survival of prostate cancer patients was analyzed using OncoMir Cancer Database (OMCD). The human prostate cancer cell line PC-3 was divided into two groups: miR-1249-5p group and negative control group. Mediated by Lipofectamine 2000, miR-1249-5p mimics liposome complex or negative miRNA liposome complex were transfected into PC-3 cell at logarithmic growth stage. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-1249-5p in PC-3 cell of two groups. Colony formation assay was used to detect the changes of the proliferation ability of PC-3 cell in the two groups. Transwell experiment was used to detect the changes of PC-3 cell invasion in the two groups, and the cell cycle changes of the two groups of PC-3 were detected by flow cytometry. The miRNA prediction software miRGator was used to predict the target gene of miR-1249-5p. RT-qPCR and Western blotting were used to detect the target gene expression of miR-1249-5p. Measurement data were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between two groups. Results:Compared with prostate cancer patients with low miR-1249-5p expression, prostate cancer patients with higher miR-1249-5p expression had longer overall survival, and the difference was statistically significant ( P<0.01). The expression level of miR-1249-5p in the miR-1249-5p group (10.74±1.19) was significantly higher than that of the negative control group (1.56±0.27), the difference was statistically significant ( P<0.01). The number of colonies formed in the miR-1249-5p group (35.86±6.94) was significantly less than that in the negative control group (88.94±11.66), and the difference was statistically significant ( P<0.01). The number of transmembrane cells [(25.01±6.83)/high power field of view] in the miR-1249-5p group was significantly less than that of the negative control group [(82.76±8.35)/high power field of view], and the difference was statistically significant ( P<0.01). The proportion of cells in the G 0-G 1 phase in the miR-1249-5p group [(50.79±6.61)%] was significantly higher than that in the negative control group [(27.09±2.30)%], the difference was statistically significant ( P<0.01), and PC-3 cell were inhibited in the G 0-G 1 phase. Neural precursor cell expressed developmentally down-regulated 9 ( NEDD9) may be the target gene of miR-1249-5p. Compared with the negative control group, the NEDD9 gene expression in the miR-1249-5p group was significantly lower than that of the negative control group, the difference was statistically significant ( P<0.01). Conclusion:miR-1249-5p can inhibit the proliferation, metastasis and cell cycle of PC-3 cell in prostate cancer, which may be achieved by negatively regulating the expression of proto-oncogene NEDD9.

Objective:To explore the relationship between the relative expression of miRNA-676-3p and the survival of renal cancer patients, and its effect on the proliferation and invasion of renal cancer by targeting and regulating prefoldin 1 (PFDN1).Methods:OncoRank online software was selected to analyze the relationship between the relative expression of miRNA-676-3p and the survival rate of renal cancer patients. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the relative expression of miRNA-676-3p in renal cancer cell lines. Renal carcinoma CAKI1 cells were resuscitated, and the transfected miRNA-NC was used as the control group, and the transfected precursor miRNA-676-3p was used as the overexpression group. The relative expression of miRNA-676-3p was detected by RT-qPCR. The cell absorbance and invasion number of the two groups were measured by CCK-8 and Transwell invasion assays, respectively. The target gene of miRNA-676-3p was predicted and verified by referring to the TargetScan Release 8.0 website and dual-luciferase reporter gene experiment. The expression of PFDN1 gene and Wnt/β-catenin molecular pathway protein in the two groups of cells were determined by RT-qPCR and Western blotting, respectively. 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 survival rate of renal cancer patients with high expression of miRNA-676-3p was significantly higher than that of renal cancer patients with low expression of miRNA-676-3p, the difference was statistically significant ( P<0.01). The relative expression of miRNA-676-3p in renal cancer cell lines was significantly lower than that in normal renal tubular epithelial cells, the difference was statistically significant ( P<0.01), and the relative expression of miRNA-676-3p in CAKI1 cells was the lowest, the difference was statistically significant ( P<0.01). The relative expression levels of miRNA-676-3p in the control and overexpression groups were 1.04±0.59 and 15.90±1.70, respectively, and the overexpression group was significantly higher than the control group, the difference was statistically significant ( P<0.01). After 24, 48, 60, and 72 h of culture, the absorbance of cells in the overexpression group was lower than that in the control group, the difference was statistically significant ( P<0.05). The number of invasion cells in the control group and the overexpression group were (115.90 ± 24.73) and (43.83 ± 21.94) cells, respectively, and the number of cell invasion in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). PFDN1 was the downstream target gene of miRNA-676-3p ( P<0.01). The relative expression of PFDN1 gene in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). The expression of Wnt/β-catenin molecular pathway proteins in the overexpression group was lower than that in the control group. Conclusions:Renal cancer patients with high expression of miRNA-676-3p had a higher survival rate. miRNA-676-3p inhibited the proliferation and invasion of renal cancer CAKI1 cells by significantly down-regulating the expression of PFDN1, thereby inhibiting the development of renal cancer.

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 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 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 explore the molecular mechanism of microRNA (miRNA, miR)-1914-3p regulating the expression of ARL4C and affecting the invasion and proliferation of renal cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of miR-1914-3p in tumor tissues and adjacent tissues of 53 renal cancer patients, 4 types of renal cancer cell lines (ACHN, OS-RC-2, 786-O, A498) and normal proximal renal tubular epithelial cell line (HK-2). The nonsense sequence (NC) and miR-1914-3p mimic were transiently transfected into renal cancer cells with the lowest miR-1914-3p expression by liposome method, namely the NC group and miR-1914-3p group. qRT-PCR was used to detect the expression level of miR-1914-3p in transfected cells. Transwell invasion test and cell counting kit-8 (CCK-8) were used to detect the invasion and proliferation ability of each group of cells. Bioinformatics software and dual luciferase gene report experiment were used to predict and test the targeted regulation mechanism of miR-1914-3p on target genes. qRT-PCR and Western blot was conducted to analyze the target gene expression level in cells of each group.Results:The expression level of miR-1914-3p in renal cancer tissue was significantly lower than that in adjacent tissues ( P<0.01). The expression level of miR-1914-3p in renal cancer cell lines was significantly lower than that in HK-2 cell lines ( P<0.01), and the expression of miR-1914-3p in OS-RC-2 cells was the lowest ( P<0.01). The expression of miR-1914-3p in the NC group and the miR-1914-3p group were (1.04±0.17) and (11.40±0.91), respectively. The expression level of miR-1914-3p in the miR-1914-3p group was significantly increased ( P<0.01), indicating that the transfection was successful. Overexpression of miR-1914-3p can significantly inhibit the invasion ( P<0.01) and proliferation ( P<0.05) of renal cancer OS-RC-2 cells. Dual luciferase gene report experiment indicated that the target gene of miR-1914-3p may be ADP-ribosylation factor-like 4C (ARL4C); miR-1914-3p can significantly inhibit the luciferase activity of wild-type ARL4C-3′UTR ( P<0.01). Overexpression of miR-1914-3p decreased the expression of ARL4C mRNA and protein in OS-RC-2 cells ( P<0.01), and decreased the expression of cell invasion phenotype proteins (Snail, Slug) and cell proliferation phenotype proteins (Mcm2, Mcm7) ( P<0.01). Conclusions:miR-1914-3p is low-expressed in renal cell carcinoma. It inhibits the invasion and proliferation of renal cell carcinoma OS-RC-2 cells through targeted interference with the expression of the oncogene ARL4C, and participates in the occurrence and development of renal cell carcinoma.