Objective To determine the transfection efficiency of recombinant adenovirus to endothelial progenitor cells(EPCs) and provide the base of lung cancer therapy by transfecting human herpes simplex virusthymidine kinase（HSV-TK） gene to EPCs. Methods Admove recombinant adenovirus 5F35(AD5F35) which transfected with βgalactosidase(AD5F35LacZ) to the 24 well plate cultivated with EPCs and transfect the EPCs. Stain the EPCs with LacZ kit and calculate the transfection efficiency. Results The blue stain cells were cells transfected successfully with AD5F35LacZ under the optical microscope. The transfection efficiencies of adenovirus to EPCs were different under the premise of the different multiplicity of infection(MOI). In a certain range, the transfection efficiencies rise with the MOI rise. When MOI was 400,the proportion of blue stain cell is the highest, which was 98.38%±1.25%. Conclusion Recombinant adenovirus can transfect EPCs successfully. The transfection efficiencies rise with the MOI rise. When the MOI is 400,the transfection efficiency is the highest.
Objective To observe endothelial progenitor cells (EPCs) participating in the formation of neovascularization in lung adenocarcinoma. Methods EPCs were transfected by recombinant adenovirus carrying LacZ gene in optimal transfection concentration, and then EPCs were injected into animal models of lung adenocarcinoma through the tail vein; afterwards, lung tissues were taken out for pathological examination in the 6th, 7th, 8th week respectively. EPCs were observed to take part in the angiogenesis in the lung adenocarcinoma through X-gal chromogenic dye. Results The optimal multiplicity of infection (MOI) of AD5F35LacZ transfected EPCs was 400. When MOI was 400, maximum transfection efficiency was 97.13±2.08. After 2 weeks, LacZ gene-transfected EPCs began to proliferate in vitro culture, then the EPCs were transplanted into animal models of lung cancer to be involved in the neovascularization formation in the 8th week after transplantation. Conclusion EPCs are involved in the formation of tumor neovascularization after transplantation.
ObjectiveTo analyse epidermal growth factor receptor (EGFR) gene mutations in pathologically confirmed lung adenocarcinoma (LAC) samples obtained by endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA). MethodsClinical data of 964 consecutive patients who underwent EBUS-TBNA in Department of Thoracic Surgery, Fudan University Shanghai Cancer Center from April 2009 to September 2013 were retrospectively reviewed. EGFR gene mutations in 77 LAC patients who were comfirmed by cell morphology and immunohistochemistry were analyzed. There were 48 males and 29 females with their median age of 61 (range 33-78) years, and 43 patients were smokers. ResultsAll the 77 LAC patients were confirmed by immunohistochemistry. Among them, 31 patients (40.26%) were found to have EGFR gene mutations. There was no statistical difference in EGFR gene mutations between male and female patients (P=0.088). Mutation rate of EGFR genes of non-smokers was significantly higher than that of smokers (P=0.032). ConclusionSamples obtained by EBUS-TBNA can be used for EGFR gene mutations analysis. The mutation rate of EGFR genes of non-smokers is higher than that of smokers.
Objective To evaluate the strategy of chemoradiotherapy following endoscopic R0 resection for esophageal cancer in M3-T1b stage. Methods There were 45 esophageal cancer patients with M3-T1b stage with endoscopic R0 resection followed by additional chemoradiotherapy from ECETC (Esophageal Cancer Endoscopic Therapy Consortium) as a trial group with 34 males and 11 females at age of 61.37±7.14 years. There were 90 patients with esophagectomy from Fudan University Shanghai Cancer Center as a control group with 63 males and 27 females at age of 61.04±8.17 years. Propensity score match (1:2) was used to balance the factors: gender, age, position, depth of invasion and lymphovascular invasion (LVI), which may influence the outcomes. Overall survival (OS) rate, relapse free survival (RFS) rate, and local recurrence rate were compared between the two groups. Result There was no statistical difference (HR=2.66 with 95%CI 0.87 to 8.11, P=0.179) in terms of OS rate between the two groups. One, two and three years overall survival rate of patients in the control group was 93%, 86%, and 84%, respectively. Nobody died in the trial group within 3 years after surgery. The RFS rate between the two groups didn’t significantly differ (HR=1.48, 95% CI 0.66 to 3.33, P=0.389). One, two and three years RFS rate of patients in the contorl group was 87%, 78%, and 76%, respectively, while 97%, 93%, and 73% in the trial group, respectively. The local recurrence rates between the two groups didn’t significantly differ either ( HR=0.53, 95%CI 0.13 to 2.18, P=0.314). One, two and three years local recurrence rate of patients in the control group was 5%, 6% and 6%, respectively, while 0%, 0% and 21% in the trial group, respectively. Conclusion Similar outcomes are found regarding OS, RFS and local recurrence rates between the two groups. The strategy of endoscopic R0 resection followed by additional chemoradiotherapy has prospect for the treatment of esophageal cancer in M3-T1b stage. And this kind of therapy may be provided for those with risk factors or can not tolerate surgery.