Long Non-coding RNA UCA1 Regulates Mitochondrial Dynamics Via MAPK Pathway To Promote The Progression Of Pancreatic Cancer

Objective:Pancreatic cancer accounts for a large proportion of cancer-related mortality in the world,and it has strong invasiveness and high metastatic potential.As a carcinogenic gene,UCA1 is involved in the occurrence and progression of a variety of tumors,including pancreatic cancer,gastric cancer,bladder cancer,colorectal cancer and so on.Abnormal mitochondrial function and metabolic changes have been recognized as important links in promoting the progression of cancer.Cancer cells change mitochondrial dynamics to resist apoptosis and adjust their biological activity and biosynthesis requirements to support tumor initiation and transformation.Previous studies have confirmed that lnc RNA affects the morphological changes of mitochondria,and UCA1 may be involved in the changes of mitochondrial dynamics in pancreatic cancer.The purpose of this study was to explore the regulation of UCA1 on mitochondrial dynamics in pancreatic cancer cells and its molecular biological mechanism in promoting the progression of pancreatic cancer.Method:1、The expression of UCA1 in pancreatic cancer cell lines(Pa Tu8988、PANC1、Bx PC-3)was detected by Real-time PCR;2、According to the basic expression level,p CDH-UCA1 plasmid was transfected into pancreatic cancer cells with lower expression of UCA1(Pa Tu8988、PANC1),and plasmid sh-UCA1 was transfected into pancreatic cancer cells with relatively high expression of UCA1(Pa Tu8988、Bx PC-3),and the transfection efficiency of each plasmid in the three kinds of cells was verified.3、The migration ability of pancreatic cancer cells was detected by Transwell migration assay,the expression of mitochondrial fusion and fission related genes was detected by Western blot,the morphological changes of mitochondria were observed under laser scanning confocal microscope,the changes of mitochondrial membrane potential were analyzed by flow cytometry,and the changes of mitochondrial autophagy were analyzed at protein level to explore the regulatory effect of UCA1 on mitochondrial dynamics in pancreatic cancer cells.4、The expression of UCA1 in pancreatic cancer cells was up-regulated or downregulated,and the difference of protein expression of ERK1/2 and JNK in MAPK pathway was detected by Western blot.Results:1、The results of Real-time PCR showed that UCA1 was differentially expressed in three kinds of pancreatic cancer cells,among which the expression of UCA1 in Bx PC-3 was the highest,the expression of UCA1 in Pa Tu8988 was relatively lower,and the expression of UCA1 in PANC1 was the lowest;2、The expression of UCA1 was successfully up-regulated in PaTu8988 and PANC1,while the expression of UCA1 was down-regulated in Bx PC-3 and Pa Tu8988;3、UCA1 promotes the progression of Pancreatic Cancer by regulating Mitochondrial Dynamics:3.1 Through Transwell assay,it was found that the migration ability of pancreatic cancer cells in the UCA1 overexpression group was stronger than that in the control group,while in the UCA1 down regulation group,the migration ability of pancreatic cancer cells was weaker than that in the control group;3.2 Flow cytometry showed that the mitochondrial membrane potential increased in pancreatic cancer cells with up-regulation of UCA1 and decreased in pancreatic cancer cells with down-regulation of UCA1;3.3 Under laser scanning confocal microscope,it was observed that the mitochondrial fusion of pancreatic cancer cells increased in the up-regulated UCA1 group;the mitochondrial fission of pancreatic cancer cells increased in the down-regulated UCA1 group;3.4 In the UCA1 overexpression group,the protein expression of mitochondrial fusion related genes increased significantly,while the fission related genes decreased,whereas in UCA1 down-regulation group,the protein expression of fission related genes increased significantly and the fusion related genes decreased;3.5 In the UCA1 overexpression group,the expression of mitochondrial autophagy related protein decreased significantly,while in the UCA1 downregulation group,the expression of mitochondrial autophagy related protein increased significantly.4、UCA1 regulates mitochondrial dynamics of pancreatic cancer cells through MAPK signal pathway:4.1 Up-regulating the expression of UCA1 in pancreatic cancer cells,Western blot assay showed that the expression of p-ERK1/2 protein related to MAPK signaling pathway was significantly increased,the expression of p-JNK protein was significantly decreased,and the expression of total ERK1/2 and JNK protein was basically unchanged;4.2 When interfering with the expression of UCA1 in pancreatic cancer cells,the expression of p-ERK1/2 related to MAPK signal pathway was significantly decreased,the expression of p-JNK was significantly increased,and the expression of total ERK1/2 and JNK was basically unchanged.Conclusion:1、UCA1 was differentially expressed in different pancreatic cancer cell lines;2、 UCA1 promotes the migration of pancreatic cancer cells and regulates the mitochondrial function of pancreatic cancer cells.Up-regulating the expression of UCA1,the mitochondrial membrane potential increases,down-regulating the expression of UCA1,the mitochondrial membrane potential decreases;up-regulating the expression of UCA1 can promote mitochondrial fusion,down-regulating UCA1 can promote mitochondrial fission;up-regulating the expression of UCA1,mitophagy is inhibited,down-regulating the expression of UCA1 mitochondrial autophagy increases;3、UCA1 can affect the mitochondrial dynamics of pancreatic cancer cells by regulating MAPK signal pathway.