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Objective To investigate the predictive and prognostic value of genetic detection of Kirsten rat sarcoma viral oncogene homolog(KRAS),Fc gamma receptors (FCGR),cytochrome P450 3A5 (CYP3A5) and CYP1A1 in patients with metastatic colorectal cancer (mCRC) receiving C225 combined with CapeOx chemotherapy.Methods Twelve KRAS wild-type (WT) mCRC patients were selected receiving C225 (cetuximab) combined with CapeOx (capecitabine/oxaliplatin) chemotherapy.KRAS,FCGR,CYP3A5,CYP1 A1 gene mutation were detected before treatment.The expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN) were detected in colorectal cancer tissues and the corresponding adjacent tissues expression with immunohistochemical staining (SP method).The relationship between FCGR,CYP3A5,CYP1A1 mutation,and PTEN expression and survival time were analyzed.Results The mutation rates of FCGR,CYP3A5,and CYP1 A1 were 16.7％ (2/12),25％ (3/12) and 16.7％ (2/12) in 12 patients with KRAS WT,respectively.PTEN was expressed mainly in the nucleus in yellowish brown.The rates of expression in the tissue adjacent to the cancer and in the tumor tissue were 100％ (12/12) and 41.7％ (5/12),respectively.PTEN expression in tumor tissue was reduced or absent.The study indicated that the objective response rate [complete response + partial response (CR + PR)] was 80％ in patients whose KRAS,FCGR,CYP3A5,and CYP1A1 were all in wild type,the CR + PR rates in FCGR,CYP3A5,and CYP1A1 gene mutation group were 0,33.6％ and 50％.The objective response rates of patients with PTEN expression or null were 50％ and 37.5％,respectively (P ＜ 0.05).The progression free survival (PFS) with KRAS/FCGR/CYP3A5/CYP1A1 wild type or mutation were 15.56 or 8.12 months (P ＜ 0.05).The overall survival (OS) with wild type or mutation of KRAS/FCGR/CYP3A5/ CYP1A1 were 25.03 or 19.21 months(P ＜0.05).The PFSs of positive or negative expression of PTEN in patients were 9.13 or 7.87 months (P ＜0.05),and the OSs of positive or negative expression of PTEN in patients were 24.25 or 18.74 months (P ＜0.05).Conclusions FCGR,CYP3A5 and CYP1A1 mutations and PTEN expression null both have been associated with resistance to cetuximab in mCRC patients.FCGR,CYP3A5,CYP1A1 and PTEN can be served as the predictive biomarkers for the response to cetuximab.
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Objective To observe the effect of PDK ( phosphoinositol -3-kinase, PI3K)/Akt-Nrf2 (Nuclear factor -E2 related factor) signal pathway on γ-glutamylcysteine synthetase (γ-GCS) in the bronchial epithelial cells of rats treated with cigarette smoke extract (CSE). Methods The bronchial epithelial cells were dealt with 10% concentration of CSE for different time and pretreated with PI3k inhibitor (LY294002). The expressions of Nrf2,p-Akt and γ-GCS proteins were examined by immunocytochemistry, flow cytometry, immunofluorescence and western blot. The expressions of γ-GCS mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR). Reduced glutathione(GSH) content and the lev-el of γ-CCS activi-ty were examined. Results GSH content in CSElh group was significantly decreased, but still higher in CSE3 and 6 groups compared to the control. Nrf2 protein mainly located in the cytoplasm. Nrf2 plasmosin mainly increased in the nucleus in control group, and Nrf2 nucleic protein significantly enhanced in CSE1, 3 and 6 groups. P-Akt protein was up-regulated at lh, reached its peak at 3h, declined slightly at 6h after exposure to CSE. The tendency of the percentage of p-Akt positive cells was as same as p-Akt protein. γ-GCS mRNA, protein and activity, gradually increased in CSE lh, CSE 3h,CSE 6h groups. Pretreated with LY294002, the expression of p-Akt protein was markedly decreases, while Nrf2 plasmosin expressed strongly, and γ-GCS mRNA, protein, activity and GSH content were significantly decreased compared to CSE3h group. Linear correlation analysis demonstrated that there were a positive correlation among Nrf2 and γ-GCS,γ-GCS activity, and among p-Akt and Nrf2,GSH,γ-GCS,γ-GCS activity. Conclusion P13K/Akt signal path might participate in Nrf2 nuclear translocation via regulating the expression of γ-GCS.
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Objective To study the expression of P73 and its relationship with expression of Survivin in colon carcinoma. Methods The expression of P73 and Survivin was studied by immunohistochemical technique in 46 cases of colon carcinoma and 12 cases of normal colon mucosa. Results The P73 expression rate of colon carcinoma was significant higher than that of normal colon mucosa(χ2 = 13.42, P ＜0.01 ). Overexpression of P73 was related to the pathological grade and Dukes stage ( ( χ2 = 8.01, P ＜0.01 ), and it was closely related to lymphatic metastasis of colon carcinoma( P ＜0.01 ). There was a positive relation between P73 and Survivin( rs =0.487, P ＜0.01 ). Conclusions The expression of P73 was closely associated with Survivin in colon carcinoma. The results suggested that the overexpression of P73 and Survivin was involved in the tumorigenesis and development of colon carcinoma and P73 and Survivin might be used as a new tumor marker in metastasis and prognosis of colon carcinoma.
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Objective To study the expression of smad2/3 protein and the effects of flunarizine on the its expression in braln tissue following transient cerebral ischemie reperfusion in gerbils. Methods A cerebral ischemia-reperfusion model in gerbils was established by clamping both common carotids. Thirty-five gerbils were randomly divided into three groups, sham operation group, cerebral ischemia-reperfusion group and flunarizine treatment group. The expression of smad2/3 protein in brain tissue was detected by immunohistochemistry technique. Results Experimental results revealed that smad2/3 protein was expressed in neuroeyte in 35 gerbil brain. Compared with sham operation group, the expression of smad2/3 protein in neurecytes of cerebral isehemia-reperfusion group was evidently increased at the lst day, 3rd day and 7th day (P <0. 01). Compared with cerebral ischemia-reperfusion group, the expression of smad2/3 protein in neurecytes of gerbils in flunarizine treatment group was evidently decreased at these time point (P < 0.05). Condusions Smad2/3 protein was expressed in nettrcvytes of gerbils. Expression of smad2/3 protein in neuroeytes of gerbils was evidently increased following cerebral ischemic reperfusion, and its expression in flunarizine treatment group was evidently decreased.
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Objective To investigate the effects of zoledronic acid on the expression of HIF-1α and VEGF in osteosarcoma LM8 cell line under hypoxic condition. Methods The hypoxic culture model was established. After LM8 cells were treated with zoledronic acid, semi-quantitative PCR was used to assess the expression of HIF-1α and VEGF mRNA. The expression of HIF-lct and VEGF protein was de-tected by immunohistochemical staining and ELISA respectively. Results Compared with cells in normoxic conditions, cells in the hypoxic environment and cells treated with zoledronic acid in the hypoxic condition did not show a significant change in the mRNA level of HIF-1α(P >0. 05). However, the protein expression of HIF-1α was markedly decreased in the cells treated with zoledronic acid in the hypoxic envi-ronment. In contrast, both mRNA and protein expression levels of VEGF were down-regulated in the zoledronic acid treatment hypoxic group (P <0.05). Conclusion Under hypoxic conditions in vitro, zoledronic acid inhibited the expression of HIF-1α protein, which decreased VEGF mRNA level and protein expression in osteosarcorna LM8 cell line.