ABSTRACT
Objective To evaluate the effect of GOLPH3 on the proliferation, apoptosis and radiosensitivity of OE33 esophageal cancer cell line. Methods The expression levels of GOLPH3 mRNA and protein in the esophageal cancer cells and normal esophageal epithelial cells were detected by qRT-PCR and Western blot, respectively. The OE33 esophageal cancer cells were transfected with GOLPH3 siRNA and subject to irradiation treatment simultaneously. The cell proliferation was detected by MTT assay. The cell apoptosis was detected by flow cytometry. The radiosensitivity was assessed by cell cloning test. The expression levels of cleaved Caspase-3, Bax and cleaved Caspase-9 protein levels were quantitatively measured by Western blot. Results The expression levels of GOLPH3 mRNA and protein in the esophageal cancer cells were significantly higher than those in the normal esophageal epithelial cells ( both P<0.05) . GOLPH3 siRNA could obviously down-regulate the expression levels of GOLPH3 mRNA and protein in the OE33 esophageal cancer cells. The proliferation activity of esophageal cancer cells was decreased, whereas the apoptosis rate was increased and the expression levels of cleaved Caspase-3, Bax and cleaved Caspase-9 were up-regulated after down-regulating the expression of GOLPH3 or irradiation treatment ( all P<0.05) . After down-regulating the expression of GOLPH3 in the esophageal cancer cells treated with irradiation, the cell proliferation activity was more significantly decreased, whereas the apoptosis rate was elevated and the expression levels of cleaved Caspase-3, Bax and cleaved Caspase-9 were more evidently up-regulated ( all P<0.05) . In the irradiated OE33 esophageal cancer cells after down-regulating the expression of GOLPH3, the radiosensitization ratio of the cells was 1.673. Conclusions GOLPH3 is highly expressed in the esophageal cancer cells. Down-regulating the expression of GOLPH3 can increase the radiosensitivity, induce the apoptosis and inhibit the proliferation of OE33 esophageal cancer cells.
ABSTRACT
BACKGROUND@#The main manifestations of radiation pneumonitis are injury of alveolar epithelial and endothelial cells, abnormal expression of cytokines, abnormal proliferation of fibroblasts and synthesis of fibrous matrix. The occurrence of radiation pneumonitis is associated with multiplecytokine level abnormality. These cytokines can also be used as bio-markers to predict the occurrence of radiation pneumonitis. This study was to evaluate the correlation between the change of apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ape1/Ref-1), intercellular adhesion molecules 1 (ICAM-1) and interleukin-17A (IL-17A) before and after radiotherapy and radiation pneumonitis for local advanced non-small cell lung cancer (NSCLC) patients with concurrent chemoradiotherapy.@*METHODS@#NSCLC patients (68 cases) were treated with concurrent radiotherapy and chemotherapy, every patient's normal tissue were controlled with a same radation dose. 68 local advanced NSCLC patients with concurrent chemoradiotherapy were detected the levels of Ape1/Ref-1, ICAM-1 and IL-17A in serum by ELISA before radiotherapy and in the 14th week after radiotherapy. Acute and advanced radiation pulmonary injury was graded according to Radiation Therapy Oncology Group/European Organization For Research and Treatment (RTOG/EORTC) diagnostic and grading criteria. Grade 2 or more radiation pneumonitis was taken as the main end point.@*RESULTS@#Eighteen cases out of 68 developed radiation pneumonitis, 50 of 68 cases have no radiation pneumonia development. There was no significant change of Ape1/Ref-1 levels before and after radiotherapy in radiation pneumonitis group (P>0.05). There was no significant change of Ape1/Ref-1 concentration in serum after radiotherapy between radiation pneumonitis group and non-radiation pneumonitis group (P>0.05). Compared with before radiotherapy, upregulation degree of ICAM-1 levels in radiation pneumonitis group was significantly higher than that in non- radiation pneumonitis group (P<0.05). There was no significant change of IL-17A concentration before and after radiotherapy in radiation pneumonitis group, but after radiotherapy IL-17A concentration in serum were remarkably higher than that in non-radiation pneumonitis group (P<0.05). Correlation analysis found that the change of ICAM-1 before and after radiotherapy has no obvious correlation with the incidence of radiation pneumonitis, and IL-17A change has obvious correlation with the incidence of radiation pneumonitis.@*CONCLUSIONS@#On the basis of strictly controlling radiation dose on normal tissue, IL-17A in serum could be the predictive factors of radiation pneumonitis for local advanced NSCLC patients with concurrent chemoradiotherapy.