Effects Of Extracts Of Cynanchum Bungei On Biological Behavior Of Human Gastric Cancer Cell Lines Regulated By LINC00858/miR-153-3P

BackgroundGastric cancer is a common malignant tumor worldwide and the third leading cause of cancer-related death in the world.So far,chemotherapy and radiation therapy are still the main treatments for patients with gastric cancer.However,radiotherapy and chemotherapy have large toxic and side effects on patients with gastric cancer,which seriously affects the quality of life of patients.Gastric cancer patients usually have resistance to radiotherapy and chemotherapy,and most patients die within one or two years.In order to reduce the mortality of gastric cancer,some new treatments are urgently needed.Chinese medicine is widely used in China and other East Asian countries.According to the theory of traditional Chinese medicine and the pathophysiology of gastric cancer,it is particularly important to develop new treatments to prevent or treat gastric cancer.Caudatin is a class of compounds extracted from the traditional Chinese medicineCynanchum bungei,which has a variety of pharmacological effects such as anti-inflammatory,antioxidant,immune regulation and anti-tumor.Recent studies have shown that Caudatin plays an antitumor role in a variety of tumors,including gastric cancer,but its molecular mechanism is not completely clear.ObjectivesThe purpose of this study was to investigate the effect of Caudatin on the biological behavior of gastric cancer cells,and to explore whether its molecular mechanism is related to the regulation of the LINC00858/miR-153-3p axis in order to provide a certain experimental basis for the treatment and prognosis of gastric cancer.Methods1.MTT assay was used to test the effects of different concentrations of Caudatin on the viability of gastric cancer AGS and HGC27 cells.Plate colony formation assay was used to test the cell clone formation ability.2.Using AnnexinV-FITC/PI cell apoptosis experiment and flow cytometry to detect the effects of different concentrations of Caudatin on the apoptosis and cell cycle of AGS and HGC27 cells.3.Transwell assay was used to detect the effects of different concentrations of Caudatin on the migration and invasion ability of AGS and HGC27 cells.4.Use qRT-PCR technology to detect the expression of lincRNA LINC00858 and miR-153-3p in gastric cancer cells and gastric mucosal epithelial cells,and the effect of Gaundatine on the expression of LINC00858 and miR-153-3p.5.The liposome transfection technology was used to transfect the LINC00858 overexpression vector into gastric cancer cells.MTT,plate clone formation,AnnexinV-FITC/PI,flow cytometry and Transwell experiments were used to detect gastric cancer cell proliferation,apoptosis,cycle,Migration and invasion capabilities.6.Predict the targeted binding sites of LINC00858 and miR-153-3p using starBase,an online database of bioinformatics,verify the targeted binding relationship between the two luciferase reporter gene experiments and Pull-down experiments,qRT-PCR detection of LINC00858 Targeting negative regulatory relationship to miR-153-3p.7.Western blot was used to detect the expression of key proteins ?-catenin,cyclinDl and c-myc in Wnt/?-catenin pathway.Results1.Caudatin can inhibit the proliferation,migration and invasion of gastric cancer AGS and HGC27 cells in a concentration-dependent manner,induce apoptosis,and block the cell cycle process.2.LINC00858 is highly expressed in gastric cancer cells,and Zaidating can inhibit the expression of LINC00858 in a concentration-dependent manner.3.Overexpression of LINC00858 can reverse the regulatory effect of Caudatin on gastric cancer cell biological behavior.4.miR-153-3p is under-expressed in gastric cancer cells,and Zadatin can up-regulate miR-153-3p expression in a concentration-dependent manner.5.LINC00858 and miR-153-3p have targeted binding sites,and LINC00858 can target the negative regulation of miR-153-3p expression.6.Caudatin can up-regulate the expression of miR-153-3p by inhibiting the expression of LINC00858.7.Caudatin can inhibit the activation of Wnt/?-catenin signaling pathway.Overexpression of LINC00858 can reverse Gatatine's inhibitory effect on Wnt/?-catenin signaling pathway.Conclusion1.Caudatin can inhibit the proliferation,migration and invasion of gastric cancer cells,induce apoptosis,and block the cell cycle process.2.Overexpression of LINC00858 can reverse the regulatory effects of Caudatin on gastric cancer cell proliferation,apoptosis,cycle,migration and invasion.3.LINC00858 can target the negative regulation of miR-153-3p expression.4.Caudatin affects the biological behavior of gastric cancer cell proliferation,apoptosis,migration and invasion by regulating the LINC00858/miR-153-3p axis.5.Overexpression of LINC00858 can reverse the inhibitory effect of Dating on Wnt/?-catenin signaling pathway.