MicroRNA-1305 Regulates The Expression Of KLF5 And The Proliferation,Invasion And Apoptosis Of Pancreatic Cancer Cells

Objective:To explore the effects of Mir-1305 on proliferation,metastasis and apoptosis of pancreatic cancer cells;The regulation of Mir-1305 and KLF5 axis on proliferation,metastasis and apoptosis of pancreatic cancer cells was explored.Method:Based on TCGA-PAAD database,The correlation between overall survival(OS)and Mir-1305,disease-specific survival(DSS)and the expression of Mir-1305,progression-free survival(PFS)and the expression of Mir-1305,and the expression of Mir-1305 before and after radiotherapy in patients with pancreatic cancer were detected.Qrt-pcr was used to detect the expression level of Mir-1305 in 26 pancreatic cancer tissues and 10 paracancer tissues,as well as the expression level of Mir-1305 in human normal pancreatic cells(HPDE6-C7 cells)and pancreatic cancer cell lines(BXPC-3,PANC-1,ASPC-1 and Pat U-8988cells).Mir-1305 was overexpressed or knocked down in BXPC-3 and PANC-1 cells by transfection with Mir-1305 mimics or inhibitors.The expression level of Mir-1305 was detected by QRT-PCR.Bxpc-3 and PANC-1 cells were transfected with Mir-1305 mimics or inhibitors,and cell viability was detected by MTT assay.Cell invasion was detected by Transwell,cell apoptosis was detected by flow cytometry,and expression levels of apoptosis-related proteins BAX,BCL2,and Cleaved caspase 3 were detected by Western Blot.Panc-1 cells infected with Agomir-1305 or Agomir-control were used to establish subcutaneous xenograft tumor models in nude mice.Real-time q PCR was used to detect the expression level of Mir-1305.The tumor volume was measured every 3 days from the 10 th day after injection,and the nude mice were sacrificed on the 25 th day to determine the tumor weight4.26 mrnas targeting Mir-1305 in pancreatic cancer were screened according to TCGA-PAAD,mi RDIP and Target Scan V7.2 databases,and the prediction of the binding sites of Mir-1305 and KLF5 was made.The targeting relationship between Mir-1305 and KLF5 was verified by luciferase reporter gene assay in 293 T cells.The effect of Mir-1305 on KLF5 expression in BXPC-3 and PANC-1 cells and the effect of overexpression of Mir-1305 on KLF5 expression in nude mouse tumor tissues were verified by QRT-PCR.The m RNA and protein expression levels of KLF5 in pancreatic cancer tissues and adjacent tissues were detected by RT-PCR and Western blot.5.Bx PC-3 and PANC-1 cells were divided into four groups by QRT-PCR: Si-NC + inhibitor NC group(cells transfected with empty plasmid),Si-KLF5 group(cells transfected with sir NA-KLF5)and Mir-1305 Expression levels of KLF5 in the inhibitor group(transfected with Mir-1305 inhibitor cells)and Si-KLF5 + Mir-1305 inhibitor or inhibitor inhibitor were tested.Cell viability was measured by MTT assay,cell invasion was measured by Transwell assay,cell apoptosis was measured by flow cytometry.Estern Blot was used to detect the expression levels of apoptosis-related proteins BAX,BCL2 and Cleaved caspase 3.Luciferase reporter gene method was used to detect the combination relationship between Mir-1305 and KLF5 in BXPC-3 and panc-1 cells.Ch IP assay was used to detect the expression relationship between KLF5 and Mir-1305 promoter in panc-1 cells.The effect of KLF5 on the expression of Mir-1305 in BXPC-3 and PANC-1 cells was analyzed by QRT-PCR.Results:Mir-1305 was down-regulated in pancreatic cancer tissue samples and cell linesMir-1305 inhibited the proliferation of pancreatic cancer cells and promoted the apoptosis of pancreatic cancer cells.The tumor weight of agomir-1305 group was significantly lower than that of agomir-NC group,and the overexpression of Mir-1305 significantly reduced the occurrence of pancreatic cancer in vivo,suggesting that the overexpression of Mir-1305 in vivo can inhibit the growth of pancreatic tumor.According to TCGA-PAAD database analysis,the expression of Mir-1305 was significantly negatively correlated with the expression of KLF5 in pancreatic cancer(R =-0.256,P = 0.00396).Mir-1305 binds to KLF5 and regulates KLF5 expression in pancreatic cancer.In cells transfected with Mir-1305 mimics,BAX and Cleaved caspase 3 were reduced,while BCL2 was significantly promoted.Mir-1305 regulates the expression level of KLF5,thereby affecting the proliferation and apoptotic progression of pancreatic cancer cells in vitro.In BXPC-3 and panc-1 cells,the lucifase activity of Mir-1305 in the Si-KLF5 group was significantly decreased,and Ch IP detection showed that KLF5 had a strong attraction to the Mir-1305 promoter in panc-1 cells.Bxpc-3 and PANC-1 cells were co-transfected with Si-KLF5 or Si-NC.The results showed that the expression level of Mir-1305 in si-KLF5 group was significantly increased.KLF5 regulates the expression of Mir-1305 as a transcriptional activator of Mir-1305 in pancreatic cancer cells.Conclusion:Mir-1305 affects the proliferation,invasion and apoptosis of pancreatic cancer cells by affecting the expression of KLF5.KLF5 binds to the promoter region of Mir-1305 and inhibits its transcription and expression.