背景:肝细胞癌(HCC)是全球癌症相关死亡的第三大原因,这在很大程度上归因于可用治疗策略的局限性。中药芪竹抗癌方(QZACP)可提高肝癌患者的生活质量,延长患者的生存时间。然而,QZACP抗癌特性的确切机制尚不清楚.
目的:本研究检查了QZACP的抗肝癌特性,特别关注其对p21激活的分泌表型(PASP)介导的免疫监视的影响,阐明肝癌的潜在分子途径。
方法:使用细胞计数试剂盒-8,5-乙炔基-2'-脱氧尿苷,和克隆检测。使用流式细胞术评估细胞周期,通过衰老相关β-半乳糖苷酶(SA-β-gal)染色鉴定衰老。在C57B/L6小鼠中建立由二乙基亚硝胺产生的原发性肝癌模型以评估QZACP的肿瘤抑制作用。采用苏木精和伊红染色检查肝脏病理特征。使用GeneCards进行PASP筛查,DisGeNet,在线孟德尔遗传在男人,和癌症基因组图谱数据库。蛋白质印迹分析,酶联免疫吸附测定(ELISA),免疫荧光染色,进行Transwell迁移测定。
结果:含QZACP的血清p21表达增强,触发细胞周期停滞,加速细胞衰老,并抑制Huh7和MHCC-97H肝癌细胞的细胞增殖。QZACP降低了肝肿瘤结节的数量和尺寸,增强了p21蛋白的表达,肿瘤病变中的SA-β-Gal染色,和细胞毒性CD8+T细胞浸润。生物信息学分析表明,PASP因素,包括肝细胞生长因子,decorin(DCN),皮肤桥蛋白,C-X-C基序趋化因子配体14(CXCL14),和Wnt家族成员2(WNT2),在肝癌的发生发展中起着重要的作用。此外,这些因素与肿瘤内自然杀伤细胞和CD8+T细胞的存在有关.Westernblotting和ELISA证实QZACP增加DCN,CXCL14和WNT2在肿瘤组织和外周血中的水平。
结论:QZACP抑制HCC进展可能涉及P21上调介导的细胞衰老,DCN,CXCL14和WNT2分泌,和逆转免疫抑制微环境。这项研究提供了可用于开发HCC新治疗策略的见解。
BACKGROUND: Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide, largely due to the limitations of available therapeutic strategies. The traditional Chinese medicine Qizhu Anticancer Prescription (QZACP) can improve the quality of life and prolong the survival time of patients with HCC. However, the precise mechanisms underlying the anti-cancer properties of QZACP remain unclear.
OBJECTIVE: This study examined the anti-hepatocarcinogenic properties of QZACP, with a specific focus on its influence on the p21-activated secretory phenotype (PASP)-mediated immune surveillance, to elucidate the underlying molecular pathways involved in HCC.
METHODS: Cell proliferation was measured using the Cell Counting Kit-8, 5-ethynyl-2\'-deoxyuridine, and clonogenic assays. The cell cycle was evaluated using flow cytometry, and senescence was identified by staining with senescence-associated beta-galactosidase (SA-β-gal). A primary liver cancer model produced by diethylnitrosamine was established in C57 BL/6 mice to assess the tumor-inhibitory effect of QZACP. The liver\'s pathological characteristics were examined using hematoxylin and eosin staining. PASP screening was performed using GeneCards, DisGeNet, Online Mendelian Inheritance in Man, and The Cancer Genome Atlas databases. Western blot analysis, enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining, and Transwell migration assays were performed.
RESULTS: Serum containing QZACP enhanced p21 expression, triggered cell cycle arrest, accelerated cell senescence, and suppressed cell proliferation in Huh7 and MHCC-97H liver cancer cells. QZACP reduced the quantity and dimensions of liver tumor nodules and enhanced p21 protein expression, SA-β-Gal staining in tumor lesions, and cytotoxic CD8+ T cell infiltration. Bioinformatic analyses indicated that PASP factors, including hepatocyte growth factor, decorin (DCN), dermatopontin, C-X-C motif chemokine ligand 14 (CXCL14), and Wnt family member 2 (WNT2), play an important role in the development of HCC. In addition, these factors are associated with the presence of natural killer cells and CD8+ T cells within tumors. Western blotting and ELISA confirmed that QZACP increased DCN, CXCL14, and WNT2 levels in tumor tissues and peripheral blood.
CONCLUSIONS: QZACP\'s suppression of HCC progression may involve cell senescence mediated via p21 upregulation, DCN, CXCL14, and WNT2 secretion, and reversal of the immunosuppressive microenvironment. This study provides insights that can be used in the development of new treatment strategies for HCC.