Chen Xuzheng, Cao Zhiyun, Zhang Youquan, Li Jinnong, Wang Suqing, Du Jian, Liao Lianming
1 Fujian University of Traditional Chinese Medicine, Fuzhou, People's Republic of China.
2 Hospital of Fuzhou University, Fuzhou, People's Republic of China.
Integr Cancer Ther. 2017 Sep;16(3):329-338. doi: 10.1177/1534735416654761. Epub 2016 Jun 22.
Fuzheng Qingjie (FZQJ) granules, a compound Chinese medicine, have been used as an adjuvant therapy for alimentary tract cancers. However, the underlying anticancer mechanisms are still not well understood. In the present study, HepG2 cells were treated with FZQJ-containing serum. Cell proliferation was evaluated using MTT assay. Apoptosis was analyzed using a flow cytometer. Cell ultrastructure was observed under a transmission electron microscope. The mitochondrial membrane potential (Δψ) was examined with JC-1 dye. In H22 tumor-bearing mice, CD4 T cells, CD8 T cells, CD3 T cells, and natural killer (NK) cells in peripheral blood were evaluated cytometrically. Interleukin (IL)-2 and tumor necrosis factor (TNF)-α levels were measured using radioimmunoassay.The mRNA levels of Bax and Bcl-2 were examined by reverse transcription-polymerase chain reaction. The protein levels of Bax, Bcl-2, cytochrome C, caspase 3 and 9, PARP, and CD69 were examined by Western blotting. The apoptotic cells in tissues were observed using TUNEL method. Alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN), and creatinine (CRE) were detected by an automatic biochemical analyzer. The results showed that FZQJ-containing serum remarkably inhibited proliferation of HepG2 cells in dose- and time-dependent manners, induced HepG2 cell apoptosis and caused a decrease of Δψ. Analysis of tumor tissue showed that FZQJ-induced apoptosis was accompanied by downregulation of Bcl-2 and upregulation of Bax, release of cytochrome c, activation of caspase 3 and 9, and cleavage of PARP. In addition, FZQJ increased the percentages of CD4 T and NK cells, the ratio of CD4/CD8 T cells as well as the levels of serum TNF-α. FZQJ also increased CD69 expression in tumor tissue. No hepatorenal toxicity was observed in H22 tumor-bearing mice. These results indicated that FZQJ could inhibit the growth of hepatoma cells via regulating immune function and inducing mitochondria mediated apoptosis.
扶正清解（FZQJ）颗粒是一种复方中药，已被用作消化道癌症的辅助治疗药物。然而，其潜在的抗癌机制仍未完全明确。在本研究中，用含FZQJ血清处理HepG2细胞。采用MTT法评估细胞增殖。使用流式细胞仪分析细胞凋亡。在透射电子显微镜下观察细胞超微结构。用JC-1染料检测线粒体膜电位（Δψ）。在荷H22肿瘤小鼠中，通过流式细胞术评估外周血中的CD4 T细胞、CD8 T细胞、CD3 T细胞和自然杀伤（NK）细胞。采用放射免疫分析法测定白细胞介素（IL）-2和肿瘤坏死因子（TNF）-α水平。通过逆转录-聚合酶链反应检测Bax和Bcl-2的mRNA水平。通过蛋白质印迹法检测Bax、Bcl-2、细胞色素C、半胱天冬酶3和9、聚（ADP-核糖）聚合酶（PARP）以及CD69的蛋白水平。采用TUNEL法观察组织中的凋亡细胞。用自动生化分析仪检测丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、血尿素氮（BUN）和肌酐（CRE）。结果表明，含FZQJ血清以剂量和时间依赖性方式显著抑制HepG2细胞增殖，诱导HepG2细胞凋亡并导致Δψ降低。肿瘤组织分析表明，FZQJ诱导的凋亡伴随着Bcl-2下调和Bax上调、细胞色素c释放、半胱天冬酶3和9激活以及PARP裂解。此外，FZQJ增加了CD4 T细胞和NK细胞的百分比、CD4/CD8 T细胞比值以及血清TNF-α水平。FZQJ还增加了肿瘤组织中CD69的表达。在荷H22肿瘤小鼠中未观察到肝肾毒性。这些结果表明，FZQJ可通过调节免疫功能和诱导线粒体介导的凋亡来抑制肝癌细胞生长。
