Abstract：
BACKGROUND: The present study aimed to elucidate the potential anticancer activity and mechanism of P. harmala\'s alkaloid extract, harmine (HAR), and harmaline (HAL) in HCT-116 colorectal cancer cells.
RESULTS: P. harmala\'s alkaloid was extracted from harmala seeds. HCT-116 cells were treated with P. harmala\'s alkaloid extract, HAR and HAL. Cytotoxicity was determined by MTT assay, apoptotic activity detected via flow cytometry and acridine orange (AO)/ethidium bromide (EB) dual staining, and cell cycle distribution analyzed with flow cytometry. The mRNA expression of Bcl-2-associated X protein (Bax) and glycogen synthase kinase-3 beta (GSK3β) was measured by real-time PCR. Furthermore, the expression of Bax, Bcl-2, GSK3β and p53 proteins, were determined by western blotting. The findings indicated that, P. harmala\'s alkaloids extract, HAR and HAL were significantly cytotoxic toward HCT116 cells after 24 and 48 h of treatment. We showed that P. harmala\'s alkaloid extract induce apoptosis and cell cycle arrest at G2 phase in the HCT116 cell line. Downregulation of GSK3β and Bcl-2 and upregulation of Bax and p53 were observed.
CONCLUSIONS: The findings of this study indicate that the P. harmala\'s alkaloid extract has anticancer activity and may be further investigated to develop future anticancer chemotherapeutic agents.
RESULTS: P. harmala\'s alkaloid was extracted from harmala seeds. HCT-116 cells were treated with P. harmala\'s alkaloid extract, HAR and HAL. Cytotoxicity was determined by MTT assay, apoptotic activity detected via flow cytometry and acridine orange (AO)/ethidium bromide (EB) dual staining, and cell cycle distribution analyzed with flow cytometry. The mRNA expression of Bcl-2-associated X protein (Bax) and glycogen synthase kinase-3 beta (GSK3β) was measured by real-time PCR. Furthermore, the expression of Bax, Bcl-2, GSK3β and p53 proteins, were determined by western blotting. The findings indicated that, P. harmala\'s alkaloids extract, HAR and HAL were significantly cytotoxic toward HCT116 cells after 24 and 48 h of treatment. We showed that P. harmala\'s alkaloid extract induce apoptosis and cell cycle arrest at G2 phase in the HCT116 cell line. Downregulation of GSK3β and Bcl-2 and upregulation of Bax and p53 were observed.
CONCLUSIONS: The findings of this study indicate that the P. harmala\'s alkaloid extract has anticancer activity and may be further investigated to develop future anticancer chemotherapeutic agents.
摘要：
背景：本研究旨在阐明P.harmala生物碱提取物的潜在抗癌活性和机制，harmine(HAR),和harmaline（HAL）在HCT-116结直肠癌细胞中的作用。
结果：P.harmala的生物碱是从harmala种子中提取的。HCT-116细胞用P.harmala生物碱提取物处理，HAR和HAL.通过MTT法测定细胞毒性，通过流式细胞术和吖啶橙（AO）/溴化乙锭（EB）双重染色检测凋亡活性，用流式细胞仪分析细胞周期分布。通过实时PCR检测Bcl-2相关X蛋白（Bax）和糖原合酶激酶3β（GSK3β）的mRNA表达。此外，Bax的表达,Bcl-2、GSK3β和p53蛋白,通过蛋白质印迹确定。调查结果表明,P.harmala生物碱提取物，治疗24和48小时后，HAR和HAL对HCT116细胞具有明显的细胞毒性。我们表明，在HCT116细胞系中，P.harmala的生物碱提取物在G2期诱导细胞凋亡和细胞周期停滞。观察到GSK3β和Bcl-2的下调以及Bax和p53的上调。
结论：本研究的结果表明，苦参生物碱提取物具有抗癌活性，有可能进一步研究开发未来的抗癌化疗药物。
结果：P.harmala的生物碱是从harmala种子中提取的。HCT-116细胞用P.harmala生物碱提取物处理，HAR和HAL.通过MTT法测定细胞毒性，通过流式细胞术和吖啶橙（AO）/溴化乙锭（EB）双重染色检测凋亡活性，用流式细胞仪分析细胞周期分布。通过实时PCR检测Bcl-2相关X蛋白（Bax）和糖原合酶激酶3β（GSK3β）的mRNA表达。此外，Bax的表达,Bcl-2、GSK3β和p53蛋白,通过蛋白质印迹确定。调查结果表明,P.harmala生物碱提取物，治疗24和48小时后，HAR和HAL对HCT116细胞具有明显的细胞毒性。我们表明，在HCT116细胞系中，P.harmala的生物碱提取物在G2期诱导细胞凋亡和细胞周期停滞。观察到GSK3β和Bcl-2的下调以及Bax和p53的上调。
结论：本研究的结果表明，苦参生物碱提取物具有抗癌活性，有可能进一步研究开发未来的抗癌化疗药物。
