1.The cell biological function and clinical significance of PRC1 in pancreatic carcinoma
Dandan Ma ; Yi Zhang ; Zhenyu Lin ; Qingtai Dong ; Zhengkang Xiao ; Zhonghu Li ; Zhiyong Zhang ; Weidong Jin
Acta Universitatis Medicinalis Anhui 2023;58(2):189-195
Objective :
To investigate the expression and prognosis of protein regulator of cytokinesis 1 ( PRC1) in pancreatic carcinoma tissues.Moreover,to explore the effects of PRC1 on the biological functions of pancreatic carcinoma cell line SW1990 and its related mechanisms.
Methods:
The GEPIA database was used to analyze the expression difference of PRC1 in pancreatic carcinoma tissues and normal pancreatic tissues.Overexpression and interference of PRC1 were achieved by Lipofectamine 3000 transfection plasmid or shRNA method.Then CCK-8 assay,Transwell assay and flow cytometry were used to detect the proliferation level,invasion ability and apoptosis of the SW1990 cells,respectively.The pancreatic carcinoma data were collected from the Cancer Genome Atlas (TCGA) database.The correlation between expression level of PRC1 and clinicopathological features of pancreatic carcinoma was analyzed.The STRING database was used to analyze the network of proteins interacting with PRC1 . Gene set enrichment analysis ( GSEA) was used to predict the possible signal pathways of PRC1 in pancreatic car- cinoma.
Results:
GEPIA database results showed that PRC1 expression in pancreatic carcinoma tissue was higher than that in normal pancreatic tissue (P<0.05) .The results of CCK-8 assay,Transwell assay and flow cytometry showed that PRC1 overexpression significantly enhanced SW1990 cell proliferation,invasion and inhibited apoptosis (P<0. 01) .Whereas PRC1 interference significantly inhibited SW1990 cell proliferation,invasion and enhanced apoptosis (P<0. 01) .TCGA database data analysis identified PRC1 mRNA expression level and M stage were independent risk factors affecting the prognosis of pancreatic carcinoma (P<0. 05) .STRING database showed that there was an interaction between PRC1 and PLK1 and so on.GSEA research results showed that the PRC1 mRNA high expression samples were enriched into P53 signaling pathway and so on (P<0. 05) .
Conclusion
PRC1 is highly expressed in pancreatic carcinoma,and it is associated with proliferation,invasion,apoptosis and prognosis of pancreatic carcinoma.Moreover,it plays an important role in pancreatic carcinoma by regulating interacting proteins PLK1 and activating P53 signaling pathways.
2.Effects of ACTL6A knockdown on proliferation , apoptosis , migration and invasion of pancreatic cancer cells
Zhenyu Lin ; Qingtai Dong ; Jianxin Zhang ; Bin Zhong ; Tao Zhang ; Zuohong Shang ; Wei Yin ; Zhonghu Li ; Dandan Ma ; Weidong Jin
Acta Universitatis Medicinalis Anhui 2022;57(10):1589-1594
Objective :
To investigate the effects of actin like 6A (ACTL6A) knockdown on the proliferation, apop⁃ tosis, migration and invasion of SW1990 cells in pancreatic cancer.
Methods :
The Oncomine database was used to analyze the expression of ACTL6A mRNA in the tissues of pancreatic cancer and normal pancreas. The plasmid of knockdown ACTL6A and siRNA negative control were established and transfected into SW1990 cell line as siRNA⁃ACTL6A group and siRNA⁃NC group. CCK⁃8, cell apoptosis experiment, Wound healing and Transwell assay were used to determine the effects of ACTL6A knockdown on the proliferation, apoptosis, migration and invasion of SW1990 cells. GSEA predicted a possible pathway regulated by ACTL6A in pancreatic cancer. T⁃test was used between the two groups.
Results :
The expression of ACTL6A in pancreatic cancer tissues was higher than that in normal pancreatic tissues ( P < 0. 05 ) . The results of CCK⁃8 assay showed that the absorbance of siRNA⁃ACTL6A group at 24 and 48 h were lower than those in the siRNA⁃NC group, and the difference was statistically significant ( t = 5. 840, 8. 454, P < 0. 01) . The results of Wound healing assay and Transwell assay showed that the healing rate and the number of invasive cells in siRNA⁃ACTL6A group were both lower than those in the siRNA⁃NC group. The difference was statistically significant ( t = 3. 960,4. 464, P < 0. 05), but the apoptosis rate of siRNA⁃ACTL6A group was significantly higher than that of the siRNA⁃NC group, and the difference was statistically significant( t = 12. 192, P < 0. 001) . GSEA results showed that the group with high expression of ACTL6A mRNA was up⁃regulated in cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway and other related gene sets(P < 0. 05) . These pathways were activated when the expression of ACTL6A was up⁃regulated.
Conclusion
ACTL6A is highly expressed in pancreatic cancer tissues. ACTL6A knockdown promotes the cell apoptosis of SW1990 cells, and inhibits proliferation, invasion and migration of SW1990 cells. The mechanism of the occurrence and development of ACTL6A in pancreatic cancer is attributed to the activation of cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway.
3.Expression of PAQR4 in hepatocellular carcinoma and its effect on the biological characteristics of HepG2 cells
Qingtai Dong ; Zhenyu Lin ; Zhonghu Li ; Zhiyong Zhang ; Dandan Ma ; Xun Cai
Acta Universitatis Medicinalis Anhui 2022;57(1):26-31
Objective :
To investigate the expression and prognosis of (progestin and adipoQ receptor family member 4 , PAQR4) in hepatocellular carcinoma (HCC) and its effect on the proliferation , invasion , migration and ap⁃
optosis of HepG2 cells.
Methods :
The HCC data in the cancer genome atlas(TCGA) database was used to analyze the expression of PAQR4 mRNA in the tissues of HCC and its prognostic significance. HepG2 cell lines of pcDNA3. 1 ⁃PAQR4 experimental group and pcDNA3. 1 ⁃vector control group were established. CCK⁃8 assay was used to detect the effect of overexpression PAQR4 on the proliferation of HepG2 cells. The scratch assay and Transwell assay were used to detect the effects of PAQR4 overexpression on the migration and invasion of HepG2 cells. PI and Annexin V double staining experiment was used to observe the effect of PAQR4 overexpression on HepG2 cell apoptosis.
Results :
TCGA database data analysis results showed that the expression of PAQR4 mRNA in HCC tissues was higher than that in adjacent tissues ( P < 0. 05 ) , and the overall survival rate of HCC patients with
PAQR4 mRNA high expression group was lower than that of low expression group (P = 0. 012) . Univariate regression analysis showed that PAQR4 mRNA expression level(HR : 1. 104 , 95% CI: 1. 051 - 1. 160 , P < 0. 001) , Tstage(HR:1.816,95% C1:1.442 -2.287,P<0.001),M stage( HR:3.924,95% CI:1.230 -12.519,P= 0. 021), pathological staging(HR:1.879, 95% C1: 1.466 -2.408, P<0. 001 ) had a significant impact on the prognosis of HCC patients. Multivariate regression analysis showed that PAQR4 mRNA expression level ( HR :1. 396 , 95% CI: 1. 081 - 1. 804 , P = 0. 011) was an independent risk factor of the prognosis of HCC patients. The results of CCK⁃8 assay , scratch assay and Transwell assay showed that the proliferation , migration , invasion and invasion ability of HepG2 cells in the experimental group were significantly improved compared with the control group (P < 0. 05) . Overexpression PAQR4 could inhibit HepG2 cell apoptosis(P < 0. 05) .
Conclusion
PAQR4 is significantly up⁃regulated in HCC tissue and is related to the prognosis of the patients. Overexpression PAQR4 promotes the proliferation , invasion and migration of HepG2 cells , and inhibits HepG2 cells apoptosis. PAQR4 maybe a new marker for the diagnosis and prognosis of HCC.