ABSTRACT
@#[Abstract] Objective: To investigate the effect of circAGFG1 on the proliferation, migration and invasion of cholangiocarcinoma QBC939 cells and its possible mechanism. Methods: The tumor tissues and corresponding para-cancerous tissues of 33 patients with cholangiocarcinoma who underwent surgical resection in the 988th Hospital of the Joint Logistics Support Force from April 2017 to October 2019 were collected. qPCR was used to detect the expression level of circAGFG1 and miR-4429 in the tissues. Cholangiocarcinoma QBC939 cells were cultured in vitro and transfected with si-circAGFG1 or miR-4429 mimics, or co-transfected with si-circAGFG1 and anti-miR-4429. Then, cell proliferation was detected by CCK-8 method and clone formation test, cell migration and invasion were detected by scratch test and Transwell assay, and the protein expression of E-cadherin and N-cadherin in cells was determined by Western blotting. Dual-luciferase reporter gene experiment was adopted to verify the regulatory relationship between circAGFG1 and miR-4429. Results: The expression of circAGFG1 was higher (3.89±0.26 vs 1.00±0.08, P<0.05) while the expression of miR-4429 (0.28±0.03 vs 1.00±0.05, P<0.05) was lower in cholangiocarcinoma tissues than those in para-cancerous tissues. After the interference with circAGFG1 or over-expression of miR-4429, the cell proliferation level, number of clone formation, scratch healing rate, number of invaded cells, and the protein expression of N-cadherin in QBC939 cells were reduced (all P<0.05), but the protein expression of E-cadherin was elevated (P<0.05). circAGFG1 could targetedly bind with miR-4429, and interfering circAGFG1 promoted the expression of miR-4429 in QBC939 cells (all P<0.05). Down-regulation of miR-4429 reversed the effect of interfering circAGFG1 on the proliferation, migration and invasion of QBC939 cells (all P<0.05). Conclusion: The expression of circAGFG1 is up-regulated in cholangiocarcinoma tissues, which may promote the proliferation, migration and invasion of cholangiocarcinoma QBC939 cells by targetedly inhibiting the expression of miR-4429.
ABSTRACT
Objective To study the effects of protein kinase C (PKC) inhibitor staurosporine (STS) on the proliferation and apoptosis of human cholangiocarcinoma QBC-939 cells and to explore its possible mechanism. Methods CCK-8 was used to detect the effects of PKC inhibitor STS on the proliferation of human cholangiocarcinoma QBC-939 cells. The effects of STS on the ultrastructural characteristics of QBC-939 cells were observed by routine transmission electron microscopy (TEM). The apoptosis rate and the cell cycle distribution of QBC-939 cells were detected by flow cytometry. The expression of cyclin B1,Cdk1 and p-Cdk1 in QBC-939 cells was detected by Western blotting. Results STS could significantly inhibit the proliferation of QBC-939 cells in a dose-dependent manner (P <0. 05) and the half inhibitory concentration ( IC50 ) of QBC-939 cells at 24th and 48th h was 334 nmol·L-1 and 118 nmol · L-1 , respectively. TEM observed that STS could induce typical apoptotic bodies and super-microstructural changes of QBC-939 cells. By Annexin V-FITC/ PI double labeling flow cytometry,we found that the apoptotic rate of QBC-939 cells after treatment with STS for 0,12,24 and 48 h was (10. 16±4. 52)% ,(22. 35±2. 19)% ,(34. 27±2. 30)% and (59. 70±5. 97)% ,respectively. By flow cytometry,compared with the control group,STS could significantly increase apoptosis rate of QBC-939 cells,decrease the percentage of cells in G0 / G1 phase and increase the percentage of cells in G2 / M phase (P<0. 05). Western blotting proved that the expression levels of cyclin B1 and Cdk1 proteins in the STS-treated QBC-939 cells were significantly decreased (P<0. 05),while the expression level of p-Cdk1 protein in the STS-treated QBC-939 cells was significantly increased ( P < 0. 05 ). Conclusion STS can significantly inhibit cell proliferation and induce apoptosis of human cholangiocarcinoma QBC-939 cells and the mechanism may be related to cell cycle arrest at G2 / M phase.
ABSTRACT
Objective To investigate the effect of hydroxycam ptothecine on PS2 and COX-2 in nude mice by human bile duct carcinoma QBC939 cells in order to analyze the correlation between PS2 and COX-2 expressions and the development of the tumor.Methods After the model of the transplanted tumor of subcutaneous tissues located the nude mice was constructed by human bile duct carcinoma QBC939 cells,30 nude mice were randomly divided into 3 groups,namely,control group,hydroxycamptothecine low-dose group and high-dose group.The growth of the transplanted tumor was observed and immunohistochemical staining combined with image analysis was used to determine the expression of PS2 and COX-2 in the tumor tissues.Results Compared with that of the control group,the content of PS2 in the tumor tissues was increased in hydroxycamptothecine low-dose group and high-dose group.The content of COX-2 was decreased and the growth of the transplanted tumor was inhibited in both hydroxycamptothecine low-dose group and high-dose group(P