| As a malignant tumor,gastric cancer?GC?originated from gastric mucosal epithelium,and its incidence rate ranks the fourth place in the world.Because the pathogenesis of the disease is still not clear,there is no very clear clinical treatment.The study reported that the pathogenic factors were associated with the imbalance between proto-oncogenes and tumor suppressor genes,which resulted in overactivation of proto-oncogenes and inhibition and inactivation of tumor suppressor genes in vivo.Gastrokine 2?GKN2?gene in tumor suppressor genes involved in the course of gastric cancer.?Janus kinase JAK1 in the body kinases1 JAK1?and JAK2?Janus kinases2,JAK2?or other cytokines to activate STAT3?signal transducers and activators of transcription3?,and thus the regulation of gastric cancer cells value-added,differentiation and apoptosis,etc.,but the specific mechanism of action of stomach cancer occur it is not clear.Curcumin?Cur?is a diketone pigment extracted from the rhizomes of some plants in the family curcumaceae and the family araceae.Studies have shown that curcumin has the effects of lowering blood lipid,anti-tumor,anti-inflammation and anti-oxidation.Curcumin may inhibit the JAK/STAT3 signaling pathway and regulate the expression of GKN2 protein,thereby affecting the proliferation and apoptosis of gastric cancer cells,thus achieving the anti-tumor effect.Objective:To study the effects of curcumin on proliferation,migration and apoptosis of human gastric cancer cell line sgc-7901 and its related mechanisms.It provides a new way for the diagnosis and treatment of gastric cancer.Methods:1.the human gastric cancer SGC-7901 cells were first resuscitated,and then the cells were cultured in RPMI 1640 medium containing 10%fetal bovine serum and 1%double antibody.Digestion and passage were performed when the cell confluence reached about 90%..Human gastric cancer SGC-7901 cells were divided into 4 groups:?1?blank control group;?2?Cur(10?mol·L-1)group;?3?Cur(20?mol·L-1)group;?4?Cur(40?mol·L-1)group;except for the blank control group,each group was added with the corresponding concentration of Cur and treated for 12h,24h,48h.The thiol blue colorimetric assay?MTT?was used to detect Cur against gastric cancer SGC-7901cells.Proliferative capacity;2.Human gastric cancer SGC-7901 cells were divided into 4 groups:?1?blank control group;?2?Cur(10?mol·L-1)group;?3?Cur(20?mol·L-1)group;?4?Cur(40?mol·L-1)group;except for the blank control group,each group was added with the corresponding drugs,and incubated for 12h,24h.The scratch test was used to detect changes in cell migration ability after drug treatment.3.Human gastric cancer SGC-7901 cells were divided into 4 groups:?1?blank control group;?2?Cur(10?mol·L-1)group;?3?Cur(20?mol·L-1)group;?4?Cur(40?mol·L-1)group;except for the blank control group,each group was added with the corresponding drugs,and incubated for 24h,48h.The apoptosis rate of curcumin treated cells was detected by Annexin-V/Pl staining flow cytometry..4.Human gastric cancer SGC-7901 cells were divided into 4 groups:?1?blank control group;?2?Cur(10?mol·L-1)group;?3?Cur(20?mol·L-1)group;?4?Cur(40?mol·L-1)Group;except for the blank control group,each group was added with the corresponding drugs and incubated for 24 hours and 48 hours.The protein expression of GKN2,JAK2,p-JAK2,STAT3 and p-STAT3 was detected by western blot.5.Human gastric cancer SGC-7901 cells were divided into 4 groups:?1?blank control group;?2?Cur(10?mol·L-1)group;?3?Cur(20?mol·L-1)group;?4?Cur(40?mol·L-1)group;except for the blank control group,the other groups were added with corresponding concentrations of curcumin and incubated for 24 hours.The protein expression of Cleaved-caspase3,Bax and Bcl-2 in human gastric cancer SGC-7901 cells was detected by western blot.Results:?1?MTT assay results:Human gastric cancer SGC-7901 cells were treated with curcumin for 12h,24h,48h,Cur(10?mol·L-1)group;Cur(20?mol·L-1)group;Cur(40?mol·L-1)group showed inhibition of proliferation.The higher the drug concentration and the longer the time,the stronger the inhibition of gastric cancer cells.?2?Scratch test showed that human gastric cancer SGC-7901 cells were treated with curcumin(10-40?mol·L-1)for 0h,12h and 24h,and the length of migration distance was detected,Cur(10?mol·L-1).Group;Cur(20?mol·L-1)group;Cur(40?mol·L-1)group showed significant migration inhibition,the higher the drug concentration and the longer the time will enhance the inhibition of gastric cancer cells.?3?Annexin-V/Pl staining flow cytometry results:human gastric cancer SGC-7901 cell line treated with curcumin for 24h and 48h(10-40?mol·L-1)was used to induce gastric cancer compared with the blank control group.Apoptosis.And the apoptotic rate of the high concentration group was significantly higher than that of the low concentration group;?4?Western Blot test results:Human gastric cancer SGC-7901 cell line treated with drug for 24 hours,human gastric cancer SGC-7901 cells treated with different concentrations of curcumin(10,20,40?mol·L-1)compared with the control group The protein expression levels of Cleaved-caspase3 and Bax were significantly increased,and the protein expression of Bcl-2 was significantly decreased.?5?Western Blot assay for GKN2 protein expression:Compared with the blank control group,the protein expression of GKN2 in curcumin-treated human gastric cancer SGC-7901 cells increased significantly with increasing concentration;curcumin-treated The relative expression levels of JAK2,p-JAK2,STAT3 and p-STAT3 proteins in human gastric cancer SGC-7901 cells were significantly lower than those in the blank control group.Conclusion:Curcumin inhibits the proliferation,migration and apoptosis of human gastric cancer SGC-7901 cells by up-regulating GKN2 protein expression and inhibiting JAK2/STAT3 pathway,thereby inhibiting the proliferation of gastric cancer cells. |
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