Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma

Satadal Das, D. Chatterjee, Banhishikha Singh, Ashoke Kumar Pradhan, Krishnendu Paira
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Abstract

Hepatocellular carcinoma (HCC) is a common cancer with high incidence rate, and 5-year survival rate in HCC is less than 20%. Thus, in search of newer anticancer agents effective in HCC, we have explored possible usefulness of an alternative medicine Lycopodium against the human liver cancer cell line, HepG2 along with its clinical efficacy. The HepG2 cell line was challenged with Lycopodium 6C (diluted Lycopodium <1pg /mL available as alternative medicine) along with vehicle alcohol control in 24 hours. The cytopathic effect and viability test with methylene blue stain were observed. The cells were harvested for total RNA extraction, and gene expression levels of targeted cytokines -Interferon gamma (IFN γ); Interleukins - IL-6, IL-8, IL-10, IL-1β, Transforming Growth Factor- TGF-β1, TGF-β3 and Tumor Necrosis Factor alpha (TNF-α) by RT-PCR were studied. DNA fragmentation assay and cell viability assay by MTT method were also tested. After ethical permission we applied this medicine as adjunct therapy to observe any beneficial role of the medicine. Statistically significant changes of IL-10, IL-1β and TGF-β3 were observed after challenge with Lycopodium 6C. The IL-10 gene expression in malignant cells was significantly reduced with Lycopodium 6C; however, the expression is more with vehicle alcohol compared to normal control set. Thus, the medicine could decrease the excessive IL-10 gene expression to a moderate level. IL-1β and TGF-β3 gene up-regulation by the vehicle alcohol were also mitigated by the medicine Lycopodium 6C. Mild DNA fragmentation was also seen in cancer cells after challenge with the medicine. Two cases suffering from hepatocellular carcinoma showed much clinical improvement after therapy with this medicine. Lycopodium 6C may act as a supporting alternative medication for treating HCC.
稀释石蒜素诱导肝癌细胞死亡及临床改善
肝细胞癌(HCC)是一种常见的高发癌症,HCC的5年生存率不足20%。因此,为了寻找对HCC有效的新型抗癌药物,我们探索了替代药物石松对人肝癌细胞系HepG2的可能有效性及其临床疗效。在HepG2细胞系24小时内,用石松6C(稀释后的石松<1pg /mL可作为替代药物)和酒精对照刺激HepG2细胞系。观察亚甲蓝染色对细胞病变的影响及细胞活力测定。收集细胞进行总RNA提取,检测目标细胞因子-干扰素γ (IFN γ)的基因表达水平;采用RT-PCR方法研究白细胞介素- IL-6、IL-8、IL-10、IL-1β、转化生长因子- TGF-β1、TGF-β3和肿瘤坏死因子α (TNF-α)的表达。采用MTT法进行DNA片段化试验和细胞活力试验。在获得伦理许可后,我们将该药作为辅助治疗来观察该药的任何有益作用。经石蒜6C攻毒后,IL-10、IL-1β、TGF-β3含量变化有统计学意义。石蒜6C可显著降低恶性细胞中IL-10基因的表达;但与正常对照组相比,载药酒精组表达更多。因此,该药物可以将过量的IL-10基因表达降低到中等水平。药石蒜6C也能减轻载体酒精对IL-1β和TGF-β3基因的上调。在接受药物攻击后,癌细胞也出现了轻微的DNA断裂。2例肝癌患者经该药治疗后临床有明显改善。石蒜6C可作为治疗HCC的辅助替代药物。
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