RNA Microarray Gene Expression Profiling Of Lung Cancer And The Function Of Metastasis And Drug Resistance With CAFs

BackgroundLung cancer had become one of the serious public health problems because of it's high incidence, progression, high recurrence and lacking of effective therapy and became one of the trouble of pulmonary diseases. Patients who had positive therapeutic response might have further invasive, lymph node metastasis and secondary drug resistant.Tumor interstitial tissue in solid tumor was a coin with two sides in the development of normal and malignant tissue. Cancer associated fibroblasts consisted the largest part of the interstitial tissue, while leading to increased matrix protein deposition, newly vascular development, and more inflammation cell infiltration. All of these changes will promote cancer cell proliferation, epithelial-mesenchymal transition and play an important role in invasion and metastasis. Still we are not clear about what had change in cancer cells with the present of CAFs. How CAFs effect lung cancer cells invasion, metastasis and drug resistant are less understood.Here, we use RNA Microarray expression profile analysis and focus on apoptosis, adhesion, drug resistant molecular for further research.Purpose1. To determine RNA Microarray expression profile difference between same cancer cell cultured with CAFs and NFs.2. To determine the reason of increased expression level of ITGB3and Bcl-2in leading cell of lung SCC.3. Weather CAFs CM could increase drug resistant of Cisplatin in lung cancer cells and the mechanism.Methods1. Primary culture of CAFs and NFs cell lines from lung tissue cut at Beijing Hospital. We detected A549cell expression profile by co-cultured with CAFs and NFs by RNA Microarray.2. Detecting Bcl-2, ITGB3expression pattern in tissue array system and compare the relationship with CAFs position. Observe Bcl-2and ITGB3expression at the present of CAFs in two kinds of co-culture system. Analysis the heterogeneous expression pattern and the relationship with clinical characteristics in lung SCC patients by x2test. 3. To verify CAFs CM can induce lung cancer cell drug resistant to Cisplatin; CAFs CM and ITGB1expression in drug resistant; In CAFs CM co-culture system to detect ITGB1down stream protein phosphorylation in drug resistant.Results1. CAFs, NFs cells were harvested by using primary culture cells and were verified.RNA Microarray found obvious change (Log2>0.5and Log2<-0.5) including85genes up-regulated and23genes down regulated in CAFs2. Lung SCC had the heterogeneous expression pattern of ITGB3and Bcl-2and they were highly expressed in adjacent to CAFs. In66cases of lung SCC tissue array cases, Bcl-2positive cases had high expression at leading cells (13/17); while ITGB3positive cases had high expression at leading cells(20/31).9in17Leading over Following cases accompanied with lymph node metastasis(81%) while negative cases was12.9%in contrast(P=0.027, x2=4.866).3. CAFs CM can induce drug resistant; CAFs CM can up-regulate ITGB1and induce drug resistant by FAK/Src-PI3K-Akt pathway.Conclusions1. This study was the first time to detect gene expression profile of lung cancer cells with the fair of CAFs and NFs from the same patient.2. This study was the first time to describe Bcl-2, ITGB3had heterogeneously expression pattern, and found this phenomenon might related to CAFs adjacent to them.3. This study was the first time to describe Bcl-2expressed in the leading edge had relation with lymph node metastasis.4. This study found CAFs can induce Cisplatin drug resistant of lung cancer by FAK/Src-PI3K-Akt pathway.