Study The Expression Of CD133 In Human Lung Adenocarcinoma Cells And The Stem Cell Properties Of Cd133-positive Human Lung Adenocarcinoma Cells

Tumor stem cells (TSC) is a small part of tumor tissue with self-renewal, unlimited proliferation and multi-differentiation capacity in tumor cells. While TSC has small proportion in the tumor tissue, it is closly related with tumor origin, development and metastasis. CD133 is a transmembrane glycoprotein found in human hematopoietic stem cells, which is thought as a surface marker of several stem cells. In 2008 Eramo for the first time reported that CD133 was expressed in the surface of lung TSC. Tirino studied 89 fresh tumor specimens of NSCLC patients with flow cytometry sorting and found there were 72% expressed CD133 in 2009. CD133-positive lung cancer cells in subsequent experiments showed more aggressive than CD133-negative cells.This experiment will detect the expression of CD133 in human lung adenocarcinoma specimens by immunohistochemistry. At the same time we will study 10 fresh human lung adenocarcinoma specimens using flow cytometry sorting and do some comparative experiments of stem cells properties to the CD133-positive and CD133-negative tumor cells.Part 1 The expression of CD133 in human lung adenocarcinoma tissues and relationship with the patients' clinical informationObjective: To study the expression of CD133 in lung adenocarcinoma specimens and to explore the relationship of CD133 expression with the patients' clinical information.Methods: To study the expression of CD133 protein in pathological paraffin specimens from selected 83 lung adenocarcinoma patients using immunohistochemistry. Doing statistical analysis combined with patient's age, sex, smoking index, lymph node metastasis, differentiation and other information, searching for the expression of CD133 in lung adenocarcinoma tissues and the correlation with patient-related information.Results: The expression rate of CD133 protein in 83 lung adenocarcinoma patients is 81.9%, at the same time the expression is heterogeneous.Conclusions: The expression of CD133 in lung adenocarcinoma is popular, but the intensity of expression is heterogeneous. The expression of CD133 is related with lymph node metastasis and the degree of pathological differentiation, but not correlated with age, sex and smoking index.Part 2 Stem cells properties study of the CD133-positive human lung adenocarcinoma cellsObjective: To get CD133-positive human lung adenocarcinoma cells by flow sorting, to assess their invasiveness, tumorigenic ability, the resistance to chemotherapy drugs and to explore whether the CD133-positive human lung adenocarcinoma cells have the tumor stem cells properties.Methods: To make 10 fresh human lung adenocarcinoma specimens into single cell suspension, to isolate CD133-positive and CD133-negative lung cancer cells after it is marked with CD133, and to do comparative study of invasion assay, the resistance to chemotherapy drug assay, immunodeficiency mice tumorigenesis assay and the expression of stem cell markers.Results: CD133 was found expressed in 8 of 10 fresh human lung adenocarcinoma specimens with the highest expression intensity was 13.12%. CD133-positive cells were much stronger than CD133-negative cells in invasiveness, tumorigenic ability in mice and the resistance to chemotherapy drugs. The expression proportion of ABCG2 in CD133-positive and CD133-negative cells have significant difference with the number of 4.61% and 2.66% respectively.Conclusions: CD133 expresses in fresh human lung adenocarcinoma specimens popularly and the expression intensity is heterogeneous. CD133-positive human lung adenocarcinoma cells compared to negative cells have more aggressive, chemotherapy-resistant and tumorigenic capacity and lung adenocarcinoma stem cells may be enriched in CD133-positive cells.