Objective:To determine the structure of triterpenoid isolated from avocado seeds and the cytotoxic effect on MCF-7 and Hep G2 cells.Methods:The powder sample was macerated with ethanol,followed with separation of the ...Objective:To determine the structure of triterpenoid isolated from avocado seeds and the cytotoxic effect on MCF-7 and Hep G2 cells.Methods:The powder sample was macerated with ethanol,followed with separation of the extract by column chromatography.The target compound was monitored on thin layer chromatography plate and reagent Lieberman–Buchard.The isolated compound was characterized by spectral analysis,mainly ultraviolet,infrared,and liquid chromatographymass spectroscopy and their spectroscopic data with those reported in literature were compared.In vitro cytotoxic activity was investigated against Vero,MCF-7,and Hep G2 cell lines using MTT assay.Results:A triterpenoid compound was isolated from ethanol extract.The extracts,fraction(F3),and the isolated compound showed a significant cytotoxic activity against all investigated cell lines.MTT assay showed that the triterpenoid isolate inhibited cell proliferation of MCF-7 and Hep G2 cell line with the IC50 values of 62 mg/m L and 12 mg/m L,respectively,and was safe to normal cells.Conclusions:The results of the present study reveal that triterpenoid from avocado seeds have the potential for further development as anticancer agents.展开更多
Background: Cancer stem cells(CSCs) accelerate the growth of hepatocellular carcinoma(HCC) residual after incomplete radiofrequency ablation(In-RFA). The present study aimed to detect the effects of In-RFA on stemness...Background: Cancer stem cells(CSCs) accelerate the growth of hepatocellular carcinoma(HCC) residual after incomplete radiofrequency ablation(In-RFA). The present study aimed to detect the effects of In-RFA on stemness transcription factors(STFs) expression which are important for the production and function of CSCs, and to find which STFs promote HCC stemness after In-RFA. Methods: HepG2 cells were used for in vitro and in vivo studies. Flow cytometry and sphere-formation assays were used to detect the level and function of CD133~+ CSCs in the models. PCR array and ELISA were applied to analyze the altered expression of 84 STFs in CD133~+ CSCs in two models. Specific lentiviral shRNA was used to knockdown STFs expression, followed by detecting In-RFA’s effects on the levels and function of CD133~+ CSCs. Results: In-RFA was identified to induce CD133~+ CSCs and increase their tumorigenesis ability in vitro and in vivo. The mRNA levels of 84 STFs in CD133~+ CSCs were detected by PCR array, showing that 15 and 22 STFs were up-regulated in two models, respectively. Meanwhile, the mRNA levels of seven common STFs were up-regulated in both models. ELISA assay demonstrated that only the protein of sex determining region Y-box 9(SOX9) was up-regulated in both models, the protein levels of the other 6 common STFs did not increase in both models. Finally, SOX9 was identified to play an important role in inducing, maintaining stemness and promoting tumorigenesis ability of CD133~+ CSCs in both models. Conclusion: In-RFA-induced SOX9 stimulates CD133~+ CSCs proliferation and increases their tumorigenesis ability, suggesting that SOX9 may be a good target for HCC treatment.展开更多
目的探讨苦参碱对肝癌HepG2细胞自噬的作用,进一步研究苦参碱诱导HepG2细胞自噬的机制。方法将0.0(对照组)、0.4、0.8、1.6g·L^(-1)苦参碱及0.4g·L~(^(-1))苦参碱+10mmol·L^(-1)自噬特异抑制剂(3-MA)分别作用于肝癌HepG...目的探讨苦参碱对肝癌HepG2细胞自噬的作用,进一步研究苦参碱诱导HepG2细胞自噬的机制。方法将0.0(对照组)、0.4、0.8、1.6g·L^(-1)苦参碱及0.4g·L~(^(-1))苦参碱+10mmol·L^(-1)自噬特异抑制剂(3-MA)分别作用于肝癌HepG2细胞,构建体外模型。采用免疫印迹法(Western blotting)检测自噬基因LC3-Ⅱ的表达,实时荧光定量PCR(Realtime-PCR)检测自噬基因WIPI1的表达,Annexin V FITC-PI流式细胞术检测HepG2细胞凋亡率,电镜观察HepG2细胞自噬体变化与凋亡情况。结果 0.4、0.8、1.6g·L^(-1)苦参碱作用的HepG2细胞LC3-Ⅱ表达相比对照组均显著增加(P<0.05),其中0.4g·L^(-1)苦参碱作用的HepG2细胞LC3-Ⅱ表达水平最高;0.4g·L^(-1)苦参碱作用的HepG2细胞自噬体、WIPI1mRNA表达水平较对照组明显增加(P<0.05)。0.4g·L^(-1)苦参碱联合3-MA作用的HepG2细胞凋亡率较单纯0.4g·L^(-1)苦参碱作用的凋亡率显著增加(P<0.05)。结论苦参碱诱导HepG2自噬且减少细胞凋亡,与WIPI1的表达相关。自噬抑制剂可抑制苦参碱诱导的自噬,促进肝癌细胞凋亡,二者联合可成为临床治疗肝癌的新措施。展开更多
基金Supported by Ministry of Finance of Indonesia through Education Fund Management Institution(LPDP)under a contract number PRJ-541/LPDP.3/2016
文摘Objective:To determine the structure of triterpenoid isolated from avocado seeds and the cytotoxic effect on MCF-7 and Hep G2 cells.Methods:The powder sample was macerated with ethanol,followed with separation of the extract by column chromatography.The target compound was monitored on thin layer chromatography plate and reagent Lieberman–Buchard.The isolated compound was characterized by spectral analysis,mainly ultraviolet,infrared,and liquid chromatographymass spectroscopy and their spectroscopic data with those reported in literature were compared.In vitro cytotoxic activity was investigated against Vero,MCF-7,and Hep G2 cell lines using MTT assay.Results:A triterpenoid compound was isolated from ethanol extract.The extracts,fraction(F3),and the isolated compound showed a significant cytotoxic activity against all investigated cell lines.MTT assay showed that the triterpenoid isolate inhibited cell proliferation of MCF-7 and Hep G2 cell line with the IC50 values of 62 mg/m L and 12 mg/m L,respectively,and was safe to normal cells.Conclusions:The results of the present study reveal that triterpenoid from avocado seeds have the potential for further development as anticancer agents.
基金supported by a grant from National Natural Science Foundation of China(81371546 and 61527807)Beijing Training Project For The Leading Talents in S&T(Z141107001514002)+4 种基金Health Industry Special Scientific Research Project(201402019)Beijing Municipal Administration of Hospitals’ Mission Plan(SML20150101)Beijing Scholar 2015(160)Capital Health Research and Development of Special Fund(2018-2-2182)the Beijing Municipal Science&Technology Commission(Z181100001718070)
文摘Background: Cancer stem cells(CSCs) accelerate the growth of hepatocellular carcinoma(HCC) residual after incomplete radiofrequency ablation(In-RFA). The present study aimed to detect the effects of In-RFA on stemness transcription factors(STFs) expression which are important for the production and function of CSCs, and to find which STFs promote HCC stemness after In-RFA. Methods: HepG2 cells were used for in vitro and in vivo studies. Flow cytometry and sphere-formation assays were used to detect the level and function of CD133~+ CSCs in the models. PCR array and ELISA were applied to analyze the altered expression of 84 STFs in CD133~+ CSCs in two models. Specific lentiviral shRNA was used to knockdown STFs expression, followed by detecting In-RFA’s effects on the levels and function of CD133~+ CSCs. Results: In-RFA was identified to induce CD133~+ CSCs and increase their tumorigenesis ability in vitro and in vivo. The mRNA levels of 84 STFs in CD133~+ CSCs were detected by PCR array, showing that 15 and 22 STFs were up-regulated in two models, respectively. Meanwhile, the mRNA levels of seven common STFs were up-regulated in both models. ELISA assay demonstrated that only the protein of sex determining region Y-box 9(SOX9) was up-regulated in both models, the protein levels of the other 6 common STFs did not increase in both models. Finally, SOX9 was identified to play an important role in inducing, maintaining stemness and promoting tumorigenesis ability of CD133~+ CSCs in both models. Conclusion: In-RFA-induced SOX9 stimulates CD133~+ CSCs proliferation and increases their tumorigenesis ability, suggesting that SOX9 may be a good target for HCC treatment.
文摘目的探讨苦参碱对肝癌HepG2细胞自噬的作用,进一步研究苦参碱诱导HepG2细胞自噬的机制。方法将0.0(对照组)、0.4、0.8、1.6g·L^(-1)苦参碱及0.4g·L~(^(-1))苦参碱+10mmol·L^(-1)自噬特异抑制剂(3-MA)分别作用于肝癌HepG2细胞,构建体外模型。采用免疫印迹法(Western blotting)检测自噬基因LC3-Ⅱ的表达,实时荧光定量PCR(Realtime-PCR)检测自噬基因WIPI1的表达,Annexin V FITC-PI流式细胞术检测HepG2细胞凋亡率,电镜观察HepG2细胞自噬体变化与凋亡情况。结果 0.4、0.8、1.6g·L^(-1)苦参碱作用的HepG2细胞LC3-Ⅱ表达相比对照组均显著增加(P<0.05),其中0.4g·L^(-1)苦参碱作用的HepG2细胞LC3-Ⅱ表达水平最高;0.4g·L^(-1)苦参碱作用的HepG2细胞自噬体、WIPI1mRNA表达水平较对照组明显增加(P<0.05)。0.4g·L^(-1)苦参碱联合3-MA作用的HepG2细胞凋亡率较单纯0.4g·L^(-1)苦参碱作用的凋亡率显著增加(P<0.05)。结论苦参碱诱导HepG2自噬且减少细胞凋亡,与WIPI1的表达相关。自噬抑制剂可抑制苦参碱诱导的自噬,促进肝癌细胞凋亡,二者联合可成为临床治疗肝癌的新措施。
