Impacts of Lobetyolin on the Proliferation,Apoptosis,and Immune Escape of Endometrial Cancer Cells by Regulating TIM3/Gal9 Signaling Pathway
Objective To investigate impacts of lobetyolin on the proliferation,apoptosis,and immune escape of endometrial cancer cells by regulating T cell immunoglobulin and mucin domain-containing molecule 3 (TIM3)/galectin-9 (Gal9)signaling pathway.Methods CCK-8 method was applied to detect the survival rate of endometrial cancer cells HEC-1-A treated with 0,15,30,60,90,120,and 150 mg/L of lobetyolin,and to further screen the optimal concentration for its action.HEC-1-A cells cultured in vitro were randomly grouped into a control group,a lobetyolin group,a TIM3 knockdown group,an empty group,and a lobetyolin+TIM3 overexpression group.After grouping and treatment, the expression of TIM3/Gal9 pathway in each group of cells was detected by real-time fluorescent quantitative PCR and western blotting;Immunofluorescence staining and CCK-8 method were applied to detect cell proliferation in each group;Cell apoptosis was detected by flow cytometry in each group;Immunofluorescence staining was applied to detect the expression of apoptosis related proteins in each group and Bax/Bcl-2 was compared.Human peripheral blood lymphocytes were co-cultured with different proportions of each group,and their cytotoxicity against each group was detected using CCK-8 method.Results Compared with the control group,expressions of TIM3,Gal9 mRNAs and proteins,the positive rate of Ki67,and the survival rate of cells in the lobetyolin group and TIM3 knockdown group all decreased (P<0.05),while the apoptosis rate,Bax/Bcl-2, and killing rate increased (P<0.05);There was no significant change in all indicators of cells in the empty group (P>0.05).Compared with the lobetyolin group,expressions of TIM3,Gal9 mRNAs and proteins, the positive rate of Ki67, and the survival rate of cells in the lobetyolin+TIM3 overexpression group all increased (P<0.05 ),while the apoptosis rate,Bax/Bcl-2,and killing rate decreased (P<0.05 ).Conclusion Lobetyolin can inhibit the proliferation and promote apoptosis of endometrial cancer cells by down-regulating the expression of TIM3/Gal9 signaling pathway,and it can also reduce their immune escape,thereby enhancing the lethality of immune cells to endometrial cancer cells.
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维普
