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Berberine enhances the anti-hepatocellular carcinoma effect of nk92-mi cells through inhibiting IFN-gamma-mediated PD-L1 expression

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Liver Research 2022年第3期6卷 167-174页

作者： Kunyuan Wang Chengxin Gu Ganxiang Yu Jiaen Lin Zhilei Wang Qianting Lu Yangzhi Xu Dan Zhao Xiaofeng Jiang Weijian Mai Shiming Liu Hui Yang Department of Gastroenterology The Second Affiliated Hospital of Guangzhou Medical UniversityGuangzhouGuangdongChina Department of Hepatobiliary Surgery The Second Affiliated Hospital of Guangzhou Medical UniversityGuangzhouGuangdongChina Guangzhou Institute of Cardiovascular Disease The Second Affiliated Hospital of Guangzhou Medical UniversityGuangzhouGuangdongChina

Background and aims:Berberine is one of the most promising clinically tested natural alkaloids,and immunotherapy using natural killer(nk)cells is a potentially effective treatment for hepatocellular carcinoma(HCC).This study aims to elucidate the effect of berberine on the anti-HCC effect of nk92-mi cells.Materials and methods:Human HCC SMMC-7721 and Hep3B cells were co-incubated with nk92-mi cells,berberine(60 or 80 mmol/L),or their combination for 36 h.To evaluate the killing effect of nk92-mi cells on HCC cells,the release of lactate dehydrogenase(LDH)in HCC cells was measured.The anti-tumor effects of berberine,nk92-mi cells,and their combinations were evaluated by MTS,EdU,Tunel,and Western blot assays.A male BALB/c nude mouse subcutaneous tumor model was used to investigate the anti-HCC effect of berberine and nk92-mi cells in vivo.Results:The LDH assay showed that berberine enhanced the cytotoxicity of nk92-mi cells on HCC cells.The combination of berberine and nk92-mi cells demonstrated more obvious anti-HCC effect than did the berberine or nk92-mi single treatment in inhibiting cell proliferation and inducing apoptosis both in vitro and in vivo.Mechanistically,the expression of programmed cell death-ligand 1(PD-L1)in HCC cells was upregulated after co-culture with nk-92mi cells.PD-L1 expression was knocked down,thereby inhibiting the proliferation and promoting apoptosis of HCC cells,and inhibited by berberine that blocked the secretion of interferon gamma(IFN-g),thereby enhancing the anti-tumor effect of nk92-mi cells.Conclusions:Current data show that the immunomodulatory role of berberine is to enhance the cytotoxic effect of nk92-mi cells and inhibit tumor immune escape by reducing the expression of PD-L1.Our study provides a rationale for the clinical application of berberine in combination with nk cells for the treatment of HCC.

关键词： Berberine Hepatocellular carcinoma(HCC) nk92-mi cells Interferon gamma(IFN-g) Programmed cell death-ligand 1(PD-L1)

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Effects of total flavonoids of Sea buckthorn (Hippophae rhamnoides L.) on cytotoxicity of nk92-mi cells

Effects of total flavonoids of Sea buckthorn (Hippophae rham...

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第十二届全国免疫学学术大会

作者：侯殿东雷萍徐铭韩晓伟关洪全辽宁中医药大学

The aim of this study was to evaluate the effects of TFH on the cytotoxicity of nk92-mi cells and its possible mechanisms.nk92-mi cells were treated with TFH(2.5 mg/L or 5.0 mg/L) or PBS for 24 h,the cytotoxicity against K562 was detected by measuring the release of lactate dehydrogenase(LDH),expression levels of NCRs and nkG2 D were detected by flow cytometry,expression levels of perforin and granzyme B were detected by Western blot.Cytokine Antibody Arrays with 80 cytokine proteins were used to profile the effect of TFH on cytokines.Western blot was adopted to detect the effects of TFH on STATl,STAT4 and STAT5 signal pathway.Compared with the normal control group,TFH could significantly enhance nk92-mi cell cytotoxicity against K562 cells,up-regulate expressions of nkp44,nkp46,perforin and granzyme B.TFH could up-regulate expressions of IL-1α,IL-2,IL-7,IL-15,CSF-2,CSF-3,MCP-1,miG,IFN-γ,TNF-α,TNF-β and down-regulate expressions of IL-16,miP-1β,CX3 CL-1 and miF.TFH could increase expressions of phospho-STAT1 and phospho-STAT5.Results demonstrated that TFH can increase nk92-mi cells cytotoxicity through STAT1 and STAT5 signal pathway.

关键词： total flavonoids of Hippophae rhamnoides L.（TFH） nk92-mi cells cytotoxicity

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