The Effect Of Sphingosine Kinase-1 On The Proliferation And Apoptosis Of SWl990 Cells And Its Expression In Pancreatic Cancer

Objective: To explore the effects of sphingosine kinase-1 on SWl990 cells and tissues of pancreatic cancer.Methods:(1)The cultured pancreatic cancer SW1990 cell lines were divided into three groups: control group,phorbol-12-myristate-13 ester group(PMA group)and dimethyl sphingosine group(DMS group).MTT method and clone formation experiment were used to test the changes of cell proliferation,flow cytometry was used to detect apoptosis,and RT-PCR method and Western blot method were used to detect the expression of sphingosine kinase-1 m RNA and protein respectively.(2)A total of 15 pairs of cancer tissues and paracancerous tissues of patients who undertook pancreatic tumor resection with pathological diagnosis of pancreatic ductal adenocarcinoma in the Department of Hepatobiliary Surgery in the First Affiliated Hospital of Hainan Medical College in 2018 were collected.And the expression of sphingosine kinase-1 m RNA and protein in pancreatic cancer tissues and paracancerous tissues were detected by semiquantitative RT-PCR and Western blot methods.Results:(1)The testing results of MTT show that the survival rate of cells in PMA group(OD value:1.37±0.03)was significantly higher than control group(OD value:1.00 ± 0.01).The survival rate of cells in DMS group(OD value : 0.65±0.02)was significantly lower than control group(P <0.05).(2)The results of clone formation show that the proliferative activity of cells in PMA group(2.12±0.18)% increased significantly compared with control group(1.39±0.27)%,and the proliferative activity of cells in DMS group(0.75 ±0.16)%decreased significantly compared with control group,both of the differences were statistically significant(P<0.05).(3)The results of flow cytometry show that the apoptosis rate was(16.71±1.53)% in control group,(9.70±0.62)% in PMA group and(31.74±1.32)% in DMS group.Statistical analysis show that the apoptosis rate of PMA group was significantly lower than control group(P<0.01),and the apoptosis rate of DMS group was significantly higher than control group(P<0.01),indicating that PMA significantly inhibits apoptosis in pancreatic cancer cells,while DMS can promote apoptosis.(4)The results of RT-PCR in three groups show that the expression of m RNA of sphingomyelin kinase-1 was 0.148 ±0.006 in control group,0.202±0.013 in PMA group and0.112±0.022 in DMS group.All the differences was statistically significant(P<0.05),indicating that the expression of sphingosine kinase-1 m RNA in PMA group was significantly higher than DMS group and control group,while the expression of sphingomyelin kinase-1 in DMS group was significantly lower than control group.(5)The results of Western blot show that the expression of sphingomyelin kinase-1protein was 0.182 ±0.044 in control group,0.258±0.015 in PMA group and0.101±0.034 in DMS group.All the differences was statistically significant(P<0.05),indicating that the expression of sphingomyelin kinase-1 protein in PMA group was significantly higher than DMS group and control group,while the expression of sphingomyelin kinase-1 protein in DMS group was significantly lower than control group.(6)The expression of sphingosine kinase-1 m RNA and protein in pancreatic cancer were significantly higher than adjacent normal pancreatic tissues(P <0.01).Conclusion: When sphingosine kinase-1 is activated,the proliferation of pancreatic cancer cells is significantly promoted,and the process of cell apoptosis is inhibited.Conversely,when sphingosine kinase-1 is inhibited,the proliferation of pancreatic cancer cells is also significantly inhibited,and cell apoptosis is promoted.In general,it can affect the proliferation and apoptosis of pancreatic cancer SWl990 cells by regulating the activity of sphingosine kinase-1.The expression levels of sphingosine kinase-1 m RNA and protein in human pancreatic cancer tissues are higher than those in normal pancreas The organization has increased significantly.Therefore,sphingosine kinase-1 plays an important role in the development of pancreatic cancer,and sphingosine kinase-1 may become an attractive target in the treatment of pancreatic cancer.