[Research of Feiliuping Gao and its combination with different types of drugs intervention on expression of PI3K/AKT/NF-κB in lung metastatic microenvironment].

The aim of this paper was to observe the effect of Feiliuping Gao and its combination with different types of drugs intervention on the expression of PI3K/AKT/NF-κB in lung metastatic microenvironment, and to reveal the advantage of Chinese medicine intervention time on the key molecule in lung metastatic microenvironment. The mouse model of Lewis lung carcinoma was established, and lung tissues were collected at 14 days, 21 days and 28 days after the intervention of Feiliuping Gao, and the expressions of PI3K, AKT and NF-κB were detected by immunohistochemistry and Western blot. At 14 days, there was no significant difference in PI3K expression between each group and the control group. The expression of AKT protein was significantly inhibited in the celecoxib (CLB) group, the Feiliuping Gao (FLP) combination with cyclophosphamide (FLP+CTX) group, and the Feiliuping Gao combination with celecoxib (FLP+CLB) group (P<0.05). The inhibition of AKT protein expression in FLP+CLB group was superior. The FLP+CLB group can inhibit the expression of NF-κB protein (P<0.05). At 21 days, compared with the control group, the expression of PI3K was inhibited in FLP group and the FLP+CTX group (P<0.05), while the expression of PI3K was best inhibited in the FLP+CLB group (P<0.001). Only the FLP+CLB group could significantly inhibit the expression of AKT protein (P<0.01). The FLP+CTX group had the best effect in inhibiting the expression of NF-κB protein (P<0.001). At 28 days, compared with the control group, the expression of PI3K and AKT was inhibited in the FLP+CLB group (P<0.001). Feiliuping ointment combination with celecoxib has an advantage in regulating the expression of PI3K/AKT/NF-κB molecules in lung metastatic microenvironment.

[Oxymatrine Diminished the Side Population and Inhibited Wnt/p-catenin Signaling Pathway in MCF- 7 Breast Cancer Cells].

Objective To observe the effects of oxymatrine (OM) in inhibiting the proliferation and percentage of cancer stem cell like cell of human breast cancer MCF-7 cells, and to study its molecular mechanism. Methods MCF-7 cells were taken as subject. Side population cells (SP) of cancer stem cell like cells rich in MCF-7 cells were isolated using side population (SP) method. The proliferation properties of SP cells and non-side population (non-SP) cells were detected by MTT assay. The proliferation and intervention of cisplatin (DDP) and OM at various concentrations were detected as well. The ex- pression levels of ß-catenin gene and protein were detected using flow cytometry and immunofluorescence technique. Results (1) OM in different concentrations had various inhibitions to the proliferation of subpopulations of MCF-7 cells. Of them, it had weakest inhibition on non-SP cells, weaker inhibition on unsorted cells, and strongest inhibition on SP cells. DDP in different concentrations had strongest inhibition on non-SP cells, weaker inhibition on unsorted cells, and weakest inhibition on SP cells. (2) SP cells accounted for 3. 1%, 1. 7%, and 0. 2% of the total cells after OM acted at 0. 25, 0. 50, 1. 00 mg/mL, respectively. The expression rate of phosphorylated ß-catenin was 42. 62% ±2. 62% after SP cells were ac- ted by OM, with statistical difference as compared with the blank control group [ (22. 8% ±1. 66%) ,P < 0. 01]. (3) Compared with SP cells without OM treatment, the expression of ß-catenin in OM treated SP cells was obviously reduced. Besides, they were specifically distributed under the cytomembrane, with nuclear translocation obviously reduced. Conclusion OM could intervene biological behaviors of cancer stem cell like cell of MCF-7 cells possibly through Inhibiting the activation of Wnt/ß-catenin signaling pathway.

Anti-tumor enhancement of Fei-Liu-Ping ointment in combination with celecoxib via cyclooxygenase-2-mediated lung metastatic inflammatory microenvironment in Lewis lung carcinoma xenograft mouse model.

BACKGROUND: Fei-Liu-Ping (FLP) ointment is an oral prescription medication that has been widely applied to treat lung cancer patients in China. Regulation of the metastatic microenvironment is an important therapeutic approach for prevention and treatment of tumor recurrence and metastasis. The advantage of Traditional Chinese Medicine management of lung cancer lies in the prevention of recurrence and metastasis. Our previous study has demonstrated that FLP ointment could regulate lung inflammatory microenvironment in vitro. However, the effects of FLP on the tumor microenvironment in vivo are still poorly understood. The objective of this study is to investigate the effect of FLP alone or in combination with celecoxib in the prevention of lung cancer progression by Cyclooxygenase (Cox)-2 mediated tumor inflammatory microenvironment in vivo. METHODS: 120 Lewis lung carcinoma xenograft mice were divided equally into four groups: vehicle, FLP, celecoxib, and FLP plus celecoxib. The dynamic growth of the xenografted tumors was observed using an in vivo fluorescence imaging system. Mice were sacrificed on day 14, day 21, and day 28, and tumor specimens and lung tissues were harvested to detect the metastasis-associated protein expression. RESULTS: Tumor inhibition rate was 15.4, 44.2, 47.4 % at day 14, 37.3, 34.7, 61.5 % at day 21, and 15.5, 10.3, 32.5 % at day 28 after treatment of FLP, celecoxib, and FLP plus celecoxib, respectively. Upon treatment of FLP and celecoxib together, lung metastasis rate was 30 % (8 metastatic nodules) lower than other groups. FLP inhibited Cox-2 expression in a time-dependent manner. Moreover, FLP inhibited N-cadherin, matrix metalloproteinases (MMP)-9, and Vimentin expression. Treatment of FLP in combination with celecoxib was more effective than FLP or celecoxib alone in inhibiting vascular endothelial growth factor, platelet-derived growth factor receptors ß, microsomal Prostaglandin E synthase-1, MMP-2, MMP-9, N-cadherin, and Vimentin expression, but increased E-cadherin expression. CONCLUSIONS: FLP inhibited tumor growth and metastasis in a Lewis lung xenograft mice model through the Cox-2 pathway. FLP in combination with celecoxib enhanced the antitumor growth and anti-metastasis effects. Traditional Chinese herbs combined with anti-inflammatory drugs might offer a promising strategy to prevent tumor metastasis.

The herb medicine formula "Yang Wei Kang Liu" improves the survival of late stage gastric cancer patients and induces the apoptosis of human gastric cancer cell line through Fas/Fas ligand and Bax/Bcl-2 pathways.

The herb medicine formula "Yang Wei Kang Liu" (YWKLF) has been used to inhibit the metastasis of human gastric cancer to prolong patient survival. In this study, we evaluated the effect of combination of chemotherapy with YWKLF on the survival of stage IV gastric cancer patients and the potential mechanisms of YWKLF by focusing on its capacity to activate apoptotic pathways in human gastric cancer cell line MGC-803. We found that combination of chemotherapy with oral administration of YWKLF significantly increased the survival of stage IV gastric cancer patients. In an approach of "serum pharmacology" in which sera were collected from rabbits orally administered with YWKLF and examined for their anti-tumor cell activity in vitro, we observed that sera from rabbits administered with YWKLF induced the apoptosis of MGC-803 cells by causing the loss of mitochondrial membrane potential, increasing the expression of Fas protein and Bax mRNA, as well as down-regulating Fas-L mRNA. Our results suggest that activation of major pro-apoptotic pathways may account for the anti-gastric cancer activity of YWKLF, which may provide a basis for isolation and identification of more highly effective anti-cancer components.

Tai Chi Chuan in postsurgical non-small cell lung cancer patients: study protocol for a randomized controlled trial.

BACKGROUND: Impairment of exercise capacity remains a common adverse effect of non-small cell lung cancer (NSCLC) survivors after surgery. Previous research has suggested that Tai Chi Chuan (TCC) offers an exercise capacity benefit in several types of cancers. This is a randomized trial to investigate the efficacy and safety of TCC in postoperative NSCLC patients over an observation period of 3 months and a 9-month follow-up. METHODS/DESIGN: Using a prospective, one center and randomized design, 120 subjects with histologically confirmed stage I-IIIA NSCLC following complete surgical resection will potentially be eligible for this trial. Following baseline assessments, eligible participants will be randomly assigned to one of two conditions: (1) TCC training, or (2) placebo control. The training sessions for both groups will last 60 min and take place three times a week for 3 months. The sessions will be supervised with target intensity of 60-80% of work capacity, dyspnea, and heart rate management. The primary study endpoint is peak oxygen consumption (VO2peak), and the secondary endpoints include: 6-min walk distance (6MWD), health-related quality of life (HRQoL), lung function, immunity function, and the state of depression and anxiety. All endpoints will be assessed at the baseline and postintervention (3 months). A follow-up period of 9 months will be included. The main time points for the evaluation of clinical efficacy and safety will be months 3, 6, 9, and 12 after enrollment. DISCUSSION: This study will assess the effect of group TCC in postsurgery NSCLC survivors on VO2peak, lung function, and other aspects. The results of this study will eventually provide clinical proof of the application of TCC as one kind of exercise training for patients across the entire NSCLC continuum, as well as information on the safety and feasibility of exercise. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR-IOR-15006548 . Registered on 12 June 2015.

Effects of Shugan Jianpi Formula () on myeloid-derived suppression cells-mediated depression breast cancer mice.

OBJECTIVE: To observe the intervention effect of Shugan Jianpi Formula (, SGJPF) on a breast cancer mouse model with depression and investigate the underlying mechanism of SGJPF in preventing the development of breast cancer. METHODS: The breast cancer model was induced by inoculation of breast cancer cells, the depression model was induced by chronic stress stimuli, and the depression cancer model was established by combining the two factors. The mice were divided into 7 groups: normal control, depression model, tumor model, depression tumor model, SGJPF, chemotherapy, and SGJPF+chemotherapy groups. The last 3 groups were depression breast cancer mice and treated respectively with SGJPF, chemotherapy drug gemcitabine (GEM), and SGJPF alongside GEM. The condition of the mice was evaluated by the expression of 5-hydroxytryptamine in hippocampus after the sucrose water test and open field test, weight change, and survival time. Tumor growth was monitored with in vivo imaging. Flow cytometry was used to analyze the level of myeloid-derived suppression cell (MDSC) in the mouse spleen, T cell subsets, and the early apoptosis of CD8+ T cells. RESULTS: The SGJPF+GEM group had the highest inhibition rate and the longest survival time (P<0.01). The MDSC level and the apoptosis rate of CD8+ T cells was the highest in the SGJPF+GEM group (P<0.05). CONCLUSIONS: Depressive disorders and tumor growth could suppress the immune function of mice to different degrees, and the microenvironment in late 4T1 inflammatory breast cancer may play an important role in the pathological process. SGJYF could regulate the immune microenvironment by reducing CD8+ T lymphocyte apoptosis and tumor cell activity, increasing immune surveillance capability, and inhibiting MDSC proliferation, thus prolonging the survival time of tumor-bearing mice.

Oxymatrine diminishes the side population and inhibits the expression of ß-catenin in MCF-7 breast cancer cells.

Cancer stem cells (CSCs) play a critical role in both cancer initiation and relapse as they are resistant to most cytotoxic agents and able to proliferate indefinitely. The plant alkaloid oxymatrine has many biological activities including the ability to induce cell cycle arrest and apoptosis, which makes it a potentially useful agent for targeting cancer cells. In order to determine whether it has beneficial pharmacological properties to eradicate CSCs, we analyzed the effects of oxymatrine on MCF-7 breast cancer cells. Cancer stem-like cells' (side population, SP) identification and sorting were performed. The inhibitory effect of oxymatrine was evaluated on the sorted SP and non-SP cells. The results indicated that oxymatrine caused a dose-dependent reduction in the proliferation of MCF-7 cells and a decrease in SP cells. Wnt/ß-catenin signaling pathway was also examined by analyzing the expression of total ß-catenin and phosphorylated ß-catenin in cytoplasm, and the results showed that the growth inhibitory effects of oxymatrine treatment on MCF-7 cells may be due to the inhibition of SP and Wnt/ß-catenin signaling pathway. Further work is warranted to explore whether oxymatrine may be a useful novel therapeutic drug for targeting breast CSCs.

Compound Kushen Injection suppresses human breast cancer stem-like cells by down-regulating the canonical Wnt/ß-catenin pathway.

BACKGROUND: Cancer stem cells (CSCs) play an important role in cancer initiation, relapse and metastasis. To date, no specific medicine has been found to target CSCs as they are resistant to most conventional therapies and proliferate indefinitely. Compound Kushen Injection (CKI) has been widely used for cancer patients with remarkable therapeutic effects in Chinese clinical settings for many years. This study focused on whether CKI could inhibit MCF-7 SP cells in vitro and in vivo. METHODS: The analysis of CKI on SP population and the main genes of Wnt signaling pathway were studied first. Then we studied the tumorigenicity of SP cells and the effects of CKI on SP cells in vivo. The mice inoculated with 10,000 SP cells were randomly divided into three groups (6 in each group) and treated with CKI, cisplatin and saline (as a control) respectively for 7 weeks. The tumor formation rates of each group were compared. The main genes and proteins of the Wnt signaling pathway were analyzed by RT-PCR and western blot. RESULTS: CKI suppressed the size of SP population (approximately 90%), and down-regulated the main genes of Wnt signaling pathway. We also determined that MCF-7 SP cells were more tumorigenic than non-SP and unsorted cells. The Wnt signaling pathway was up-regulated in tumors derived from SP cells compared with that in tumors from non-SP cells. The tumor formation rate of the CKI Group was 33% (2/6, P < 0.05), and that of Cisplatin Group was 50%(3/6, P < 0.05), whereas that of the Control Group was 100% (6/6).The RT-PCR and western blot results indicated that CKI suppressed tumor growth by down-regulating the Wnt/ß-catenin pathway, while cisplatin activated the Wnt/ß-catenin pathway and might spare SP cells. CONCLUSIONS: It suggested that CKI may serve as a novel drug targeting cancer stem-like cells, though further studies are recommended.

Protective effects of tetrahydropalmatine against gamma-radiation induced damage to human endothelial cells.

AIMS: Irradiation-induced damage to pulmonary endothelial cells is thought to be an important mediator of the pathogenesis of radiation pneumonopathy. Tetrahydropalmatine (THP) has been shown to have a protective effect against oxidative stress. This study was designed to investigate the potential radioprotective effect of THP against irradiation-induced endothelial cellular damage and to elucidate the underlying mechanisms. MAIN METHODS: Human EA.hy926 cells were treated with THP and irradiation. Cell viability was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For the detection of apoptosis, morphological observation, flow cytometry and a caspase-3 activity assay were employed. The expression of cytochrome-c and Bax/Bcl-2 protein were detected by western blot analysis. Generation of reactive oxygen species (ROS) was measured by flow cytometry. Malondialdehyde (MDA), lactate dehydrogenase (LDH), glutathione (GSH) and superoxide dismutase (SOD) were measured to assess cellular oxidative stress induced injury. KEY FINDINGS: Preincubation of EA.hy926 cells with THP before gamma-radiation resulted in significant inhibition of apoptosis and enhancement of cell viability, as revealed by morphological observation, flow cytometry and MTT assay. THP significantly reduced intracellular ROS formation, levels of intracellular MDA and LDH, and enhanced the production of intracellular antioxidants (GSH and SOD) in EA.hy926 cells. Meanwhile, THP also inhibited the decrease of intracellular mitochondrial membrane potential (psim), caspase-3 activation, cytochrome-c release and reduced Bax/Bcl-2 ratio in THP pretreated, irradiated cells. SIGNIFICANCE: Our findings demonstrated THP could effectively protect endothelial cells against gamma-irradiation injury, which could potentially be applied to the prevention of endothelial cell dysfunctions associated with ionizing irradiation-induced lung injury.