ABSTRACT
Objective To evaluate the role of circular RNA protein arginine methyltransferase 5 (circPRMT5) in the genesis and progression of colorectal cancer.Methods From January 2013 to December 2017,96 patients with colorectal cancer who underwent radical resection in Department of General Surgery,Jiading District Central Hospital Affiliated to Shanghai Medical College of Health were collected.The expression of circPRMT5 in colorectal cancer tissues was examined by real-time polymerase chain reaction (RT-PCR).The correlation between circPRMT5 expression level and age,gender,tumor size,tumor location,pathological differentiation,TNM stage,lymph node metastasis of patients with colorectal cancer was analyzed.The SW620 and LOVO cells were divided into control group,circPRMT5-lenti group and circPRMT5-shRNA-lenti group according to different interventions.The effects of circPRMT5 expression level on viability,apoptosis,mitochondrial membrane potential and migration of SW620 and LOVO cells were detected.The influence of circPRMT5 expression level on E-cadherin,Slug,N-cadherin and vimentin was determined by Western blotting method.The potential target miRNA of circPRMT5 was predicted by Starbase V2.0.Student's t test,analysis of variance and chi-square test were performed for statistical analysis.Results The results of RT-PCR showed that the expression of circPRMT5 in colorectal cancer tissues was higher than that of adjacent cancer tissues (2.167 ± 0.345 vs.1.103 ± 0.144),and the difference was statistically significant (t =26.847,P ＜ 0.01).The circPRMT5 expression level was positively correlated with tumor size,TNM stage,lymph node metastasis and distant metastasis (x2 =6.010,10.971,5.321 and 6.272,all P ＜0.05).The upregulation of circPRMT5 expression could promote proliferation and migration of SW620 and LOVO cells.The circPRMT5 downregulation could inhibit cell proliferation,induce apoptosis and decrease mitochondrial membrane potential.The results of Western blotting indicated that,compared with those of control group,the expression of Slug,N-cadherin and vimentin increased in circPRMT5-1enti group (1.023 ±0.038 vs.2.105 ±0.042,1.051 ±0.309 vs.2.277 ± 0.111,1.055 ± 0.040 vs.2.002 ± 0.537,respectively),however the expression of E-cadherin decreased (2.074 ± 0.214 vs.0.627 ± 0.023),and the differences were statistically significant (t =31.817,22.065,14.536 and 9.148,all P ＜ 0.01).Compared with the control group,the expression of Slug,N-cadherin and vimentin decreased in circPRMT5-shRNA-lenti group (1.023 ± 0.038 vs.0.585 ± 0.023,1.051 ± 0.309 vs.0.616 ± 0.043,1.055 ±0.040 vs.0.537 ±0.022),while the expression of E-cadherin increased (2.074 ± 0.214 vs.2.756 ± 0.148),and the differences were statistically significant (t =-13.795,-14.252,-11.794 and-13.116,all P ＜ 0.05).A total of 21 miRNAs might have potential binding sites with circPRMT5 predicted by Starbase V2.0 software.The expression of miRNA4735-3p,miRNA202-3p,miRNA326,let-7i-5p and miRNA4500 was negatively correlated with circPRMT5 expression in both SW620 and LOVO cells confirmed by RT-PCR.Conclusion CircPRM75 is an important oneogenic gene in the genesis and progress of colorectal cancer,and may have certain potential application prospect in the research and development for colorectal cancer.
ABSTRACT
Objective To investigate the effects of IL-1 β on proliferation and migration of gallbladder cancer cells.Methods The secretion of IL-1 β in tissues of gallbladder cancer, chronic cholecystitis and normal gallbladder as well as in supernatant of gallbladder cancer cell lines (GBC-SD, SGC996) and HIBEpic cells was determined by enzyme-linked immunosorbent assay (ELISA) method.The levels of IL-1 β mRNA in GBC-SD, SGC996 and HIBEpic cells were measured by RT-PCR assay.The effects of exogenous IL-1 β on the proliferation of GBC-SD and SGC996 cells in vitro and in vivo were evaluated using WST-1 assay and xenograft tumor model, respectively.The effects of exogenous IL-1 β on the migration of GBC-SD and SGC996 cells in vitro were measured by Tranwell assay.The levels of Twist protein in GBC-SD and SGC996 ceils were examined by western blot assay after treatment with exogenous IL-1 β.In addition, the proliferation and migration of GBC-SD and SGC996 cells after gene silencing of Twist by Twist-siRNA were also evaluated.Results The level of IL-1β protein in normal gallbladder was low (66.4 ± 35.0)pg/ml,while it was significantly increased in gallbladder cancer and chronic cholecystitis [(616.4 ± 95.7) pg/ml and (422.3 ± 48.9) pg/ml, P ＜ 0.05].The levels of IL-1 βin GBC-SD and SGC996 cell culture medium [(587.4 ± 99.8) pg/ml and (657.2 ± 76.6) pg/ml] were much higher than those in the HIBEpic cells [(38.4± 12.1)pg/ml, P ＜ 0.05].Exogenous IL-1β promoted the proliferation of GBC-SD and SGC996 cells both in vitro and in vivo as well as migration in vitro (P ＜ 0.05).The level of Twist protein was significantly increased after treatment with exogenous IL-1 β.In addition, gene silencing of Twist blocked IL-1 β-induced proliferation and migration of GBC-SD and SGC996 cells.Conclusion IL-1 β promoted proliferation and migration of gallbladder cancer cells via Twist activation.