| Objective: Liver cancer is one of the most common life-threatening cancer.More than 50% of the world's new cases and deaths patients with liver cancer occurr in China.Liver cancer ranks the second cancer mortality in all cancers in China,among which hepatocellular carcinoma(HCC)accounts for 85% to 90% of liver cancer.The previous studies found that HCC are associated with hepatitis,nonalcoholic fatty liver disease(NAFLD)and alcoholic fatty liver(AFLD),and the prognosis is poor.Sterol regulatory element binding protein 1(SREBP1)is an important transcriptional factor in the biosynthesis of cholesterol,fatty acids and phospholipids.The amount of evidence also indicates abnormal expression of SREBP1 in a variety of metabolic diseases,such as diabetes,NAFLD/NASH(Non-alcoholic steatohepatitis),pathological obesity,hyperlipidemia and so on.SREBP1 is exceptionally expressed in a variety of tumors and is closely related to the degree of malignancy of the tumor.Polygoni multiflori Thunb(Heshouwu)belongs to Polygonaceae which is continuously documented in the different versions of the Chinese Pharmacopeia.Its processed form,Polygoni multiflori Radix Praeparata(Zhiheshouwu),is bitter,sweet and astringent in taste,and slightly warm in flavor.Zhiheshouwu can supplement the liver and kidney,nurish the essence and blood,blacken the beard and hair,strengthen sinew and bone,resolve turbidity and reduce lipid.Polygoni multiflori Radix Praeparata has anti-hyperlipidemia and anti-tumor effects.However,it remains a matter if inhibiting cancer cells is related to the effects of lowering lipid metabolism.In this study,the relationship between the two and the underlying mechanisms were investigated.Methods:(1)Zhiheshouwu ethanol extract liquid(HSWE)was prepared and the bioactive components were determined by high performance liquid chromatography(HPLC)according to the criterion of the Chinese Pharmacopeia(Version 2015);(2)Human hepatocellular carcinoma Bel-7402 cells were divided into lipid-normal and-deficient groups and the experiments were conducted as following:(1)Cell growth of Bel-7402 cells with HSWE(1000,600,400,250,125,25 and 0 ?g/mL)in the lipid-normal/deficient media in 12 and 24 h was observed by OD value of MTT assay.(2)JC-10 was used to determine the alteration of mitochondrial membrane potential;Cell apoptosis was determined by Hoechst 33258 staining and flow cytometry(FCM);Samples were collected to determine the content of total TG,TC,ALT and AST by Biochemimical analyzer;The level of olieic acid,stearic acid,palmitic acid and palmitoleic acid was measured by UPLC-MS;The transcriptomics change in HSWE-treated HCC cells was sequenced and analyzed;The mRNA level of SREBP1,SCD1,FABP3,ACACA and ACLY was measured by RT-PCR;The protein level of SREBP1,SCD1 and FABP3 was detected by western blot.Results:(1)The results by HPLC detection showed that the concentration of TSG and the free anthranquinones(calculated by emodin plus physcion)in HSWE was 8.09% and 0.10%,respectively;(2)MTT assay showed that cell growth of Bel-7402 cells with lipid-normal/deficient media were inhibited by HSWE in 12 and 24 h.After 12 h HSWE treatment,the level of alanine transaminase(ALT)and aspartate transaminase(AST)in Bel-7402 cells were increased.The mitochondrial membrane potential(??m)was decreased.HSWE induced Bel-7402 cells apoptosis.(3)HSWE could decrease the level of TG and TC in Bel-7402 cells,the ratio of olieic acid/ stearic acid and palmitoleic acid/ palmitic acid were decreased.(4)The western blot and RT-PCR results showed that HSWE inhibited lipogenesis protein and mRNA expression including SCD1 and FABP3 in Bel-7402 cells.(5)The data showed that after treated with HSWE for 12 h,the expression of SREBP1 mRNA and protein of the activated SREBP1 was decreased.Conclusion: HSWE induces an intrinsic apoptosis by bloking SREBP1-mediated transcriptional regulation in hepatocellular carcinoma cells. |
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