背景:肝细胞癌(HCC)是全球范围内具有高发病率和死亡率的侵袭性杀手。草本植物Chloranthusfortunei(A.灰色)Solms-Laub被称为“四季风”,在经典瑶药中被归类为冯型药。根据姚明的医学信念,金花具有驱风功能,调节气,解毒,促进血液循环,等。民间用它的汤剂来治疗停滞的肝脏疾病,如肝脓肿,肝硬化,肝炎,还有肝癌.然而,青草提取物抗肝癌的生物活性和机制尚未报道。
目的:研究五味子提取物(CFS)的抗肝癌生物活性和潜在机制。
方法:使用70%乙醇回流提取CFS,得到CFS乙醇提取物,然后用石油醚依次萃取,氯仿,乙酸乙酯,和正丁醇,产生四个部分。CCK-8测定用于检查4个部分对MHCC97-H和HepG2细胞的细胞毒性作用。探索最有效的组件,即CFS的石油醚提取物(PECFS)。采用LC/MS技术对PECFS的主要活性成分进行鉴定,并研究了肝癌细胞对细胞增殖和凋亡的影响。使用RT-PCR和Western印迹验证了该途径中的关键基因和蛋白。选择BALB/c裸鼠进行肿瘤异种移植和PECFS治疗。抑制肝癌细胞增殖,促进细胞凋亡。
结果:在四个部分中,结果表明,PECFS对MHCC97-H和HepG2细胞具有最高的抗增殖活性(IC50=13.86,10.55μg/mL),倍半萜化合物是主要的活性成分。PECFS对HCC细胞的抗增殖活性与细胞克隆的抑制有关,入侵,和转移能力,以及细胞周期停滞在G2/M期。此外,通过上调促凋亡蛋白Bax对肝癌细胞发挥促凋亡作用,下调抗凋亡蛋白Bcl-2,并激活Caspase家族的表达。此外,蛋白和m-RNA表达数据显示,PECFS通过调节PI3K/AKT/mTOR通路抑制HCC细胞增殖,促进细胞凋亡。此外,经过PECFS治疗,裸鼠的肿瘤生长受到抑制。
结论:PECFS可以通过作用于PI3K/AKT/mTOR通路抑制肝癌细胞的活力,证明了抗肿瘤的潜力。这项研究的结果表明,PECFS可能代表肝癌治疗新药物的一个有希望的来源,为CFS治疗HCC的传统应用提供科学依据。
BACKGROUND: Hepatocellular Carcinoma (HCC) is an aggressive killer worldwide with high incidence and mortality. The herb Chloranthus fortunei (A. Gray) Solms-Laub is known as \"Si Ji Feng\" and is classified as a Feng-type medicine in classic Yao medicines. According to Yao\'s medical beliefs, Chloranthus fortunei has the functions of dispelling Feng, regulating qi, detoxifying, promoting blood circulation, etc. Folk uses its decoctions to treat stagnant liver conditions, such as liver abscesses, cirrhosis, hepatitis, and liver cancer. However, the bioactivity and mechanisms of Chloranthus fortunei extract against HCC have not been reported.
OBJECTIVE: To investigate the anti-HCC bioactivity and potential mechanism of the extract of Chloranthus fortunei (CFS).
METHODS: Using 70% ethanol for reflux extraction of CFS resulted in the CFS ethanol extract, followed by sequential extractions with petroleum ether, chloroform, ethyl acetate, and n-butanol, yielding four fractions. The CCK-8 assay was utilized to examine the cytotoxic effects of 4 fractions on MHCC97-H and HepG2 cells, exploring the most effective component, namely petroleum ether extracts of CFS (PECFS). The major active ingredients of PECFS were identified using LC/MS technology, and the impact on cell proliferation and apoptosis in HCC cells was studied. The key genes and proteins in the pathway were validated using RT-PCR and Western blotting. BALB/c nude mice were chosen for tumor xenotransplantation and PECFS therapy. hinders the proliferation of HCC cells and promotes apoptosis.
RESULTS: Among the four fractions, it was found that PECFS have the highest antiproliferative activity against MHCC97-H and HepG2 cells (IC50 = 13.86, 10.55 μg/mL), with sesquiterpene compounds being the primary active constituents. The antiproliferative activity of PECFS on HCC cells was linked to the inhibition of cell cloning, invasion, and metastasis abilities, as well as the arrest of the cell cycle at the G2/M phase. Additionally, exerts pro-apoptotic effects on HCC cells by upregulating the pro-apoptotic protein Bax, downregulating the anti-apoptotic protein Bcl-2, and activating the expression of the Caspase family. Moreover, protein and m-RNA expression data showed that PECFS inhibits HCC cell proliferation and promotes apoptosis by regulating the PI3K/AKT/mTOR pathway. Besides, after PECFS treatment, tumor growth in nude mice was suppressed.
CONCLUSIONS: PECFS can inhibit the viability of HCC cells by acting on the PI3K/AKT/mTOR pathway, demonstrating anti-tumor potential. This study\'s findings suggest that PECFS may represent a promising source of novel agents for liver cancer treatment, providing scientific evidence for the traditional application of CFS in treating HCC.