The Research Of Relationship Between Nano Silica Stimulating BEAS-2B Cells-induced Inflammation And Fibrotic Processes And Peripheral Blood-related Cytokines

Background and Objective:Lung cancer is a malignant tumor occurred in the bronchial epithelium. The etiology of lung cancer has not been discovered. Lung cancer outbreaks highly in Yunnan province, especially in Xuanwei. Female patients in Xuanwei have the highest lung cancer morbidity in the world. Previous researches have shown that the high morbidity of lung cancer may be related to residence used-coal which contains large amount of nano silicon dioxide particles. If people inhale the smoke from these particles, their lung will be inflamed.There are four factors participating in lung inflammation: ① Promoting tumor factors:IL-6, IL-8, IL-17, RANTES and MIP-1a;②Antitumor factors:IL-4, IL-13, IL-2 and TNF α-and TNFγ;③Factors promoting angiogenesis and fibrosis:VEGF, b-FGF, MCP-1, IL-12 and IL-5. ④There is a special factor IL-10, which has two sides influence on the development of tumor. It can be conducive to the survival of tumor or inhibiting to its growth. This research will use different particle size of nanometer silicon dioxide to stimulate normal human bronchial epithelial cells (BEAS-2B cells) and discuss their deep relationship. Furthermore, this research will explore the role of inflammation in the lung cancer occurrence and progress to lay the foundation for the future development of proper tumor markers.Methodology:1. This research will use the vitro cell culture method, adding different particle size of nanometer silicon dioxide into 1640 medium. This method will stimulate normal human bronchial epithelial cells (BEAS-2B cells) through different particle size of nanometer. The morphological changes (size and structure) will be observed in 24 hours. Concentrations of the silicon dioxide are 10μg/mL、25μg/mL and 50ug/mL. After 24 hours, the supernatant liquid will be extracted after centrifugation.2. This research will take advantage of the Bio-Plex suspension chip system to detect the changes in factors after mentioned above when BEAS-2B cells are stimulated by nano silicon dioxide for 24 hours. These changes includes ① Promoting tumor factors:IL-6, IL-8, IL-17, RANTES and MIP-1a; ②Antitumor factors:IL-4, IL-13, IL-2 and TNF-a and TNF-γ; ③Factors promoting angiogenesis and fibrosis:VEGF, b-FGF, MCP-1, IL-12 and IL-5. ④The special factor IL-10 which has two sides influence on the development of tumor.3. This research will collect samples from 20 female NSCLC patients in Xuanwei and 20 female NSCLC patients outside of Xuanwei and peripheral blood samples from 20 normal female. The peripheral blood samples will be taken from the upper serum after centrifugal, according to different clinical stage, pathological type, the degree of differentiation and lymph node metastasis. The Bio-Plex suspension chip system will detect serum tumor type factors: ① Promoting tumor factors:IL-6, IL-8, IL-17, RANTES and MIP-1a;②Antitumor factors:IL-4, IL-13, IL-2 and TNF-a and TNF-γ; ③Factors promoting angiogenesis and fibrosis:VEGF, b-FGF, MCP-1, IL-12 and IL-5. ④The special factor IL-10 which has two sides influence on the development of tumor.Results:1. Morphology change of BEAS-2B under the stimulation of cell nanometer silicon dioxide:after 24 hours, compared with the control group, experimental group grow poorly when seen in the microscopic observation. Cells grow up in size. Some cells can be seen in the cytoplasm vacuoles change. Their edges are dull and irregular. Besides, some cells appear to float under 50ug/mL concentration.2. Nano silicon dioxide works detersively on BEAS-2B cell inflammatory reaction. In different concentrations, different size of nano silicon dioxide can cause different changes in the BEAS-2B cells factors in the supernatant fluid. Antitumor class factor: The RANTES factors differs between in 30 nm and 50 nm groups (P<0.05). Promote angiogenesis and fibrosis factor:VEGF factor differs before the control group and the 30 nm group (P<0.05). IL-12 factor differs between the control group,30 nm and 50 nm group (P<0.05). The IL-10 factor differs all in the control group,30 nm and 50 nm group (P<0.05).3. Results from detecting the peripheral blood condition of xuanwei female lung cancer patients is shown below. The amount of Anti-tumor class factor is different in female patients from Xuanwei, non-Xuanwei and normal female. IL-6 factors differs significantly between the xuanwei group and normal group (P<0.01), IL17 factor diverse in the normal group and the Xuanwei group. It also diverse in the normal group and the non-Xuanwei group. (P<0.05). Moreover, antitumor factor changes as well. IL-4 factors differs between normal group and the xuanwei group (P<0.05). There were significant differences of IL-13 between normal group and Xuanwei and non-Xuanwei group (P<0.01). INF-y differs between the Xuanwei group and normal group (P<0.05). Moreover, results can be make from the fibrosis factor promoting angiogenesis. VEGF factor differs hugely between normal group and Xuanwei,, non-Xuanwei group (P<0.01). B-FGF factor differs between normal group and the Xuanwei (P<0.05). IL-12 factor also differs hugely between normal group and Xuanwei group, non-Xuanwei group (P<0.01); Promoting tumor and antitumor factor IL-10 differs between the Xuanwei group and normal group (P<0.05) and differs largely between the normal group and the Xuanwei group (P<0.01).Conclusion:1. Nano silicon dioxide has stimulus action effect on the bronchial epithelial cells. It can promote the bronchial epithelium in severe atypical hyperplasia, which may be associated with lung cancer.2. To a certain extent, the patient condition can be detected by examining inflammation and fibrosis of related cytokines secreted state. Tumor markers with great clinical significance can be promoted in the future.3. Xuanwei women patients with lung cancer have different expression factors, which may berelated to the high incidence of lung cancer of female in Xuanwei, Yunnan Province.