RÉSUMÉ
Objective To construct the prokaryotic expression vector of human esophageal cancer related gene 4 (ECRG4), to purify the recombinant ECRG4 protein and to verify the biological function of the recombinant ECRG4 protein. Methods DNA recombination technology was utilized to construct the ECRG4 protein prokaryotic expression vector. The recombinant ECRG4 protein was purified with the transformation of escherichia coli. Then the purity of the recombinant ECRG4 protein was examined by using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Furthermore, esophageal cancer EC-18 cell line was treated by recombinant ECRG4 protein (10 μg/ml) for phosphate buffer (PBS) 48 h respectively, and the tumor cell cycle change was examined by using flow cytometry. Results SDS-PAGE analysis showed that the high purity of recombinant ECRG4 protein was obtained and the ECRG4 protein prokaryotic expression vector was successfully constructed. The cell proportion of G1 phase in PBS control group was lower than that in the recombinant ECRG4 protein group [(60.4 ±2.1) ％ vs. (71.6 ±1.8) ％; t= 25.695, P= 0.002]. The cell proportion of S phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(24.6±1.4) ％ vs. (16.5±1.0) ％; t= 36.905, P= 0.001]. The cell proportion of G2/M phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(15.0 ±1.1) ％ vs. (11.9 ±0.8) ％; t=6.471, P=0.023], which indicated that the recombinant ECRG4 protein could induce the G1 phase arrest of EC-18 cells. Conclusion The ECRG4 protein prokaryotic expression vector is successfully constructed. And the recombinant ECRG4 protein has an active biological function in esophageal carcinoma.
RÉSUMÉ
Objective To investigate the expression profile of cancer-related genes in human bone marrow-derived neural stem cells (Md-NSCs) to determine whether there are any characteristics that could help the evaluation of their tumorigenic potentials.Methods Md-NSCs were cultured in vitro and identified (experimental group);fresh human adult bone marrow cells were used as control group (sifting erythrocytes).The expression profiles of 440 cancer-related genes in cells from the two groups were analyzed by Oligo GEArray Human Cancer Microarray OHS-802;real-time quantitative PCR was performed to detect the expressions of oncogene MYC,matrix metalloproteinase 2 (MMP2),Notch congener 2 (Notch2),stanniocalcin 1 (STC1),integrin α3 (ITGA 3),signal transduction and transcriptional activation factor 5b (STA T5b),Ras congene gene family C (RhoC),and wingless-type MMTV integration site family member 1 (Wnt1).Results As compared with those in the control group,the Md-NSCs from experimental group had 66 tumor-related genes with high expressions (＞3 folds).MYC,MMP2,Notch2,STCI,ITGA3,STA T5b,RhoC and Wnt1 expressions in the Md-NSCs from experimental group were significantly higher than those in the control group (P＜0.05),whose results were accorded with genechip detection results,enjoying the folds of 4.35×100,2.84×100,2.87×100,3.41 ×102,2.22×102,6.99× 100,4.92 × 100 and 3.64 ×100,respectively.Conclusion A number of cancer-related genes are over-expressed in Md-NSCs,and the activations of some of these important oncogenes have been proved to promote human tumorigenesis.
RÉSUMÉ
Objective To investigate the expression situation of thyroid cancer related gene-1 (TC-1),CyclinD1 andβ-catenin in the tissues of non-small cell lung cancer(NSCLC)and their relationship with the clinical pathologic characteristics,to analyze their relationship with the regulation of Wnt/β-catenin signal pathway to provide the basis for studying the role of TC-1 in NSCLC. Methods The expressions of TC-1,CyclinD1 and β-catenin in 48 patients with NSCLC were detected by immunohistochemical SP method and analyzed by combining the clinical pathological features.Results The expression levels of TC-1,CyclinD1 andβ-catenin in the NSCLC tissue were significantly higher than those in the normal lung tissue;in which,the expression of TC-1 in NSCLC tis-sue was associated with lymph node metastasis and TNM stages;the expression ofβ-catenin in NSCLC tissue was related with the pathological types and tissue differentiation degree.Conclusion The expressions of TC-1,CyclinD1 andβ-catenin show the up-regu-lating trend in NSCLC and may play an important role in the development of lung cancer;TC-1 may be involved in the regulation of Wnt/β-catenin signaling pathway,which provide the new research thought of the NSCLC targeted therapy.