Objective: To evaluate the anticancer potentials of Annona muricata fruit by in vitro and in vivo methods. Methods: The ethanolic extract of Annona muricata fruit was prepared by Soxhlet extraction method and further ...Objective: To evaluate the anticancer potentials of Annona muricata fruit by in vitro and in vivo methods. Methods: The ethanolic extract of Annona muricata fruit was prepared by Soxhlet extraction method and further fractionated with petroleum ether, ethyl acetate and chloroform. The fractions were tested for cytotoxicity, apoptosis, scratch wound assay, and cell cycle analysis. IC50, apoptotic index and percentage cell migration were determined using HepG2 cells. For the in vivo studies, hepatocellular carcinoma was induced by administering 0.01% diethylnitrosamine(DEN) in drinking water in Wistar rats. In pre-treatment, rats were co-administered 200 mg/kg of fruit extract with DEN for 14 weeks. In post-treatment, the extract was co-administered after 8-weeks of DEN-induction for 14 weeks. Liver function test, haematological test, oxidative stress markers, relative liver weight, number of cancer nodules and histopathological parameters were determined.Results: Annona muricata fruit extract =significantly lowered cell proliferation counts. The chloroform-fraction possessed higher activity (IC50=(53.7±4.3) μg/mL)The chloroform fraction inhibited cell migration, which was significant compared to curcumin. Further investigations regarding the mode of anticancer activity revealed that the chloroform fraction induced apoptosis. The cell cycle analysis indicated that cells were being arrested at G0/G1. In the in vivo studies, the DEN-control group showed a significant decrease in body weights with increased mortality rate, hepatic nodules, and impairment of liver function compared to normal rats. The rats pre-treated and post-treated with the extract showed positive results with significant improvement in the parameters that were adversely affected by DEN. In addition, other adverse effects of DEN, such as blood dyscrasias and hepatic endogenous antioxidant, were significantly attenuated by Annona muricata fruit extract.Conclusions: The Annona muricata fruit extract has anticancer activity when tested by in vitro and in vivo hepatocellular cancer models.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
文摘Objective: To evaluate the anticancer potentials of Annona muricata fruit by in vitro and in vivo methods. Methods: The ethanolic extract of Annona muricata fruit was prepared by Soxhlet extraction method and further fractionated with petroleum ether, ethyl acetate and chloroform. The fractions were tested for cytotoxicity, apoptosis, scratch wound assay, and cell cycle analysis. IC50, apoptotic index and percentage cell migration were determined using HepG2 cells. For the in vivo studies, hepatocellular carcinoma was induced by administering 0.01% diethylnitrosamine(DEN) in drinking water in Wistar rats. In pre-treatment, rats were co-administered 200 mg/kg of fruit extract with DEN for 14 weeks. In post-treatment, the extract was co-administered after 8-weeks of DEN-induction for 14 weeks. Liver function test, haematological test, oxidative stress markers, relative liver weight, number of cancer nodules and histopathological parameters were determined.Results: Annona muricata fruit extract =significantly lowered cell proliferation counts. The chloroform-fraction possessed higher activity (IC50=(53.7±4.3) μg/mL)The chloroform fraction inhibited cell migration, which was significant compared to curcumin. Further investigations regarding the mode of anticancer activity revealed that the chloroform fraction induced apoptosis. The cell cycle analysis indicated that cells were being arrested at G0/G1. In the in vivo studies, the DEN-control group showed a significant decrease in body weights with increased mortality rate, hepatic nodules, and impairment of liver function compared to normal rats. The rats pre-treated and post-treated with the extract showed positive results with significant improvement in the parameters that were adversely affected by DEN. In addition, other adverse effects of DEN, such as blood dyscrasias and hepatic endogenous antioxidant, were significantly attenuated by Annona muricata fruit extract.Conclusions: The Annona muricata fruit extract has anticancer activity when tested by in vitro and in vivo hepatocellular cancer models.
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。
