Feiyanning Formula Induces Apoptosis of Lung Adenocarcinoma Cells by Activating the Mitochondrial Pathway.

Feiyanning formula (FYN) is a traditional Chinese medicine (TCM) prescription used for more than 20 years in the treatment of lung cancer. FYN is composed of Astragalus membranaceus, Polygonatum sibiricum, Atractylodes macrocephala, Cornus officinalis, Paris polyphylla, and Polistes olivaceous, etc. All of them have been proved to have anti-tumor effect. In this study, we used the TCM network pharmacological analysis to perform the collection of compound and disease target, the prediction of compound target and biological signal and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. It was found that the activation of mitochondrial pathway might be the molecular mechanism of the anti-lung cancer effect of FYN. The experimental results showed that FYN had an inhibitory effect on the growth of lung cancer cells in a dose-dependent and time-dependent manner. Moreover, FYN induced G2/M cell cycle arrest and apoptotic cell death as early as 6 h after treatment. In addition, FYN significantly induced mitochondrial membrane depolarization and increased calreticulin expression. Metabolomics analysis showed the increase of ATP utilization (assessed by a significant increase of the AMP/ATP and ADP/ATP ratio, necessary for apoptosis induction) and decrease of polyamines (that reflects growth potential). Taken together, our study suggested that FYN induced apoptosis of lung adenocarcinoma cells by promoting metabolism and changing the mitochondrial membrane potential, further supporting the validity of network pharmacological prediction.

Clinical observation of Shuanghuang Shengbai Granule () on prevention and treatment of myelosuppression caused by chemotherapy in cancer patients.

OBJECTIVE: To study the efficacy and safety of Shuanghuang Shengbai Granule (, SSG), a traditional Chinese herbal medicine, on myelosuppression of cancer patients caused by chemotherapy. METHODS: A total of 330 patients were randomly assigned to the treatment group (220 cases, analysed 209 cases) and the control group (110 cases, analysed 102 cases) with a 2:1 ratio by envelope method. The patients in the treatment group at the first day of chemotherapy started to take SSG for 14 days, while the patients in the control group took Leucogon Tablets. The changes of the blood routine, clinical symptoms and immune function in both groups were observed for safety and efficacy evaluation. RESULTS: At the 7th day of chemotherapy, the white blood cells (WBCs) level in the treatment group was significantly higher than that in the control group (P<0.05). After treatment, the WBCs rate in the normal range accounted for 50.2% in the treatment group, the myelosuppression of WBCs and neutrophil were mainly grade I, while 8.1% and 5.7% of patients emerged grade III and grade IV myelosuppression, respectively. The incidence of myelosuppression of the treatment group was significantly lower than that of the control group (P<0.05). The total effective rate of Chinese medicine syndrome in the treatment group was significantly higher than that in the control group (84.2% vs. 72.5%, P<0.05). The immune cell levels in both groups were maintained in the normal range. Compared with that before treatment, the levels of CD3+ and CD4+ cells were significantly increased in the treatment group after treatment (P<0.05). The discrepancy of CD3+ and CD4+ cell activity before and after treatment in both groups were significantly different (P<0.05). No obvious adverse event occurred in both groups. CONCLUSION: SSG had a protection effect on bone marrow suppression, and alleviated the clinical symptoms together with clinical safety.

[Effect of Shuanghuang Shengbai Granule on Wnt Signal Transduction Pathway in Tumor-bearing Mice with Chemotherapy Induced Myelosuppression].

Objective To observe the regulation of Shuanghuang Shengbai Granule (SHSBG) on regulating Wnt signaling pathway in tumor-bearing mice with chemotherapy induced myelosuppression. Methods Chemotherapy induced myelosuppression model was established in Lewis lung tumor bearing mice by intraperitoneal injection of cyclophosphamide (CTX). And then they were intervened by SHSBG. Routine white blood cell (WBC) count, red blood cell (RBC) count, platelet count, and tumor mass were calculated. Ratios of bone marrow hematopoietic stem cell (Sca, CD34 double positive cells) were detec- ted by flow cytometry. mRNA expression of main genes in Wnt signaling pathway (Wnt, ß-catenin, Frizzted, DSH, GSK3) were detected using real time fluorescent quantitative PCR. Results The number of WBC and ratio of hematopoietic stem cells in the treatment group were higher than those in the model group (P<0. 05). Expressions of Wnt, ß-catenin, Frizzted, DSH, and GSK3 mRNA in the bone marrow were higher in the treatment group than in the model group (P <0. 05). Expressions of Wnt, ß-catenin, Frizzted, and DSH mRNA expression in tumors were lower in the treatment group than in the model group (P <0. 05). There was no statistical difference in counts of RBC and platelet, tumor mass, or GSK3 mR- NA expression among all groups (P >0. 05). Conclusions The mechanism for SHSBG treating myelo-suppression was related to regulating Wnt signaling pathway. Besides, it had dual regulation effect on Wnt signaling pathway, up-regulating expressions of main genes in Wnt signaling pathway while inhibiting ex- pressions of partial genes in tumors.

[Effect of bufalin combined gefitinib on lung cancer H1975 cells and its mechanisms research].

OBJECTIVE: To observe the effect of bufalin combined Gefitinib on lung cancer H1975 cells, and to explore its potential mechanisms for anti-tumor. METHODS: The cytostatic effects of bufalin (1 -100 nmol/L), gefitinib (0.1-20 micromol/L), and bufalin plus gefitinib on H1975 cells were evaluated by MTT assay. Their effects on apoptosis of H1975 cells were determined by flow cytometry (FCM). Their effects on expressions of epidermal growth factor receptor (EGFR) and Met signal pathway related proteins in H1975 cells were detected by Western blot. RESULTS: Results of MTT assay showed that gefitinib over 5 micromol/L could inhibit H1975 cells. But combined therapy of bufalin and gefitinib could potently inhibit the growth of H1975 cells. Results of FCM showed the apoptotic rate was 61.64% +/- 5.61% in the bufalin plus gefitinib group, obviously higher than that of the bufalin group (18.34% +/- 3.42%) and the gefitinib group (7.32% +/- 1.08%), showing statistical difference (P < 0.01). Results of Western blot showed the protein expressions of p-EGFR, p-Met, p-Akt, and p-mTOR in H1975 cells could be markedly down-regulated by bufalin plus gefitinib. CONCLUSIONS: Combination of bufalin and gefitinib potently inhibited the growth of H1975 cells, and induced cell apoptosis. The potential mechanism for anti-tumor might be involved in blocking EGFR-PI3k/Akt pathway.

[The relationships between angiogenesis and matrix metalloproteinase-2, -9 in mice tumor intervened by feiyanning recipe's components].

OBJECTIVE: To observe the effects of Feiyanning Recipe's (FR) components on the microvessel density (MVD), the mRNA and protein expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 in Lewis tumors loaded in C57BL/6 mice. METHODS: C57BL/6 Lewis lung cancer mouse model was established. Mice were randomly divided into five groups, i.e., the model group, the FR group, the qi benefiting group, the Shen-tonifying group, and the anti-cancer group. The mice were killed on the 22nd day after 21-day gastrogavage. Tumors were extracted. The MVD of Lewis tumor was detected by immunohistochemical SP method. The mRNA and protein expressions of MMP-2 and MMP-9 were measured using reverse transcription-polymerase chain reaction (RT-PCR) and SP method. RESULTS: Compared with the model group and the qi benefiting group, the MVD was significantly reduced in the FR group, the Shen-tonifying group, and the anti-cancer group (P<0.01, P<0.05). But there was no significant difference between the qi benefiting group and the model group (P>0.05). The mRNA and protein expressions of MMP-2 in the FR group, the Shen-tonifying group, and the anti-cancer group were also significantly less than those in the model group (P<0.05, P<0.01). At the same time the expression of MMP-2 mRNA in the Shen-tonifying group and the anti-cancer group was also less than that in the qi benefiting group (P<0.05). The mRNA and protein expressions of MMP-9 in the FR group, the Shen-tonifying group, and the anti-cancer group were significantly lower than those in the model group and the qi benefiting group (P<0.05, P<0.01). CONCLUSION: The target for Shen-tonifying and anti-cancer Chinese herbs to inhibit tumor angiogenesis might be correlated with restraining expressions of MMP-2 and MMP-9, and protecting tumor microenvironment.

[Effects of chemotherapy combined with Chinese herbal medicine Kangliu Zengxiao decoction on tumor markers of patients with advanced non-small-cell lung cancer: a randomized, controlled trial].

BACKGROUND: Tumor markers are widely used in clinical practice and have become important indicators in assessing cancer progress. There is increasing concern that chemotherapy combined with traditional Chinese medicine has effects in decreasing the level of tumor markers. OBJECTIVE: To investigate the effects of chemotherapy combined with Kangliu Zengxiao Decoction (KLZX), a compound Chinese herbal drug, on tumor markers carbohydrate antigen 50 (CA 50), cytokeratin 19 fragment (CYFRA21-1) and carcinoembryonic antigen (CEA) in patients with advanced non-small-cell lung cancer (NSCLC) and to explore the relationships between clinical efficacy and tumor markers. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: Patients were included from Punan Hospital of Shanghai Pudong New District and Longhua Hospital between October 2008 and December 2009. Seventy-four subjects with advanced NSCLC were randomly assigned into treatment group (n=37) and control group (n=37). Patients in the control group were treated with chemotherapy alone while patients in the treatment group were treated with chemotherapy combined with KLZX. Chemotherapy of NP (vinorelbine + cisplatin) was given for two cycles and patients in the treatment group were administered with KLZX during chemotherapy. MAIN OUTCOME MEASURES: Levels of CA50, CYFRA21-1 and CEA before and after treatment were evaluated and the relationship between changes in levels of tumor makers and tumor size, clinical symptoms and living condition score (Karnofsky score) was analyzed. RESULTS: No patients achieved a complete remission. The disease control rates (complete remission (CR)+partial remission (PR)+no change (NC)) were 89.20% (33/37) and 70.30% (26/37) in the treatment and control group respectively (P<0.05). The levels of CA50, CYFRA21-1 and CEA were clearly decreased in the treatment group after treatment (P<0.05) while also decreased in the patients without progression of disease. There were no obvious changes of CA50, CYFRA21-1 and CEA in the control group, and there was even a trend of increase. Furthermore, the improvement rates of clinical syndrome were 51% (19/37) vs 11% (4/37) (P<0.05) in the treatment group and control group respectively. The total response rates of quality of life were 91.89% (34/37) vs 56.76% (21/37) (P<0.01) in the treatment and control group respectively. CONCLUSION: Combined chemotherapy with KLZX in treating advanced NSCLC can acquire better stabilizing tumor foci, decrease levels of tumor markers and improve the clinical symptoms and Karnofsky score.

Treatment of advanced non-small-cell lung cancer with Chinese herbal medicine by stages combined with chemotherapy.

PURPOSE: To observe the effect of Chinese herbal medicine by stages combined with chemotherapy in treating patients with non-small-cell lung cancer (NSCLC) of stage III or IV. METHODS: Adopting prospective, randomized, controlled, multi-centered trial design, 121 patients enrolled were assigned to the treatment group (n = 65) and the control group (n = 56). All the patients were randomized to receive chemotherapy alone or chemotherapy and Chinese herbal medicine combined (Kangliuzengxiao decoction during chemotherapy and Feiyanning decoction after chemotherapy). The main outcome measures were survival time, Karnofsky score, main clinical symptoms, and adverse reactions. RESULTS: Five patients discontinued from the trial due to oral administration of Iressa after disease progression or other reasons, and 116 patients were evaluable for clinical efficacy with 63 in the treatment group and 53 in the control group. The overall response rate were 15.87% vs. 7.55% (P = 0.170), and the disease control rate were 85.71% vs. 71.70% in the treatment and control group (P = 0.063), respectively. The median survival time was 16.17 months vs. 12.00 months in the treatment and control group (P = 0.165), respectively. In addition, adverse reactions such as leucopenia in the treatment group were less than those in the control group (P = 0.039). CONCLUSIONS: Chinese herbal medicine combined with chemotherapy showed favorable effect in improving quality of life and prolonging survival time on patients with advanced NSCLC.

[Inhibitory effects of Feiyanning Decoction on proliferation, migration and tube formation of human umbilical vein endothelial cells in vitro].

OBJECTIVE: To observe the inhibitory effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine for replenishing qi, nourishing essence, and diminishing stagnation by detoxification, on proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were isolated from the new-born umbilical core of human. Serum containing Feiyanning was prepared in SD rats by oral gavage of the Feiyanning Decoction for three days. The inhibitory effects of different contents of serum containing Feiyanning on the proliferation of HUVECs, human lung adenocarcinoma cell A549, and HUVECs cultured with A549-conditioned media were observed by sulforhodamine B (SRB) method; the effects on migration of HUVECs were inspected by using Boyden Chamber Transwell method, and the effects on tube formation of HUVECs were evaluated by cavity forming experiment. RESULTS: Serum containing Feiyanning at a concentration of 25%, 37.5% or 50% inhibited the proliferation of both the HUVECs and the HUVECs cultured with A549-conditioned media (P < 0.01, P < 0.05). High concentration serum containing Feiyanning inhibited the proliferation of A549 cells. Sera containing Feiyanning at concentrations of 12.5% to 37.5% inhibited the migration of HUVECs induced by 20% fetal bovine serum (P < 0.01, P < 0.05) and the formation of the primary tube of HUVECs. CONCLUSION: Feiyanning Decoction can inhibit the angiogenesis of HUVECs, which may be one of its mechanisms in inhibiting the invasion metastases of lung cancer.