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BACKGROUND:Pax6 gene plays an important role in eye development and differentiation, and to study how it regulates the differentiation of human bone marrow mesenchymal stem cel s (BMSCs), gaining the BMSCs stably over-expressing Pax6 is crucial, which is also the basis of stem cel replacement therapy. OBJECTIVE:To construct a lentivirus vector containing Pax6 and detect the expression of Pax6 in transfected human BMSCs. METHODS:Pax6 gene was extracted using PCR. After its connection with lentivirus vector pHIV-EGFP, it was then packaged by 293T cel s. The human BMSCs were transfected with recombinant lentivirus Pax6-EGFP as wel as lentivirus vector pHIV-EGFP, which was considered negative control group. The cel ular morphology was observed by a fluorescence microscope, and the mRNA expression of Pax6 was detected by real-time PCR. RESULTS AND CONCLUSION:The recombinant lentivirus Pax6-EGFP was constructed successful y with a titer of 3×109 pfu/L. After the transfection, both the green fluorescent protein and Pax6 gene were expressed detected using fluorescence microscope and real-time PCR, showing that the method of lentiviral transfection is a safe and effective way to modify BMSCs.
ABSTRACT
Purpose To investigate the expression of C-C chemokine receptor 9 (CCR9) in non-small cell lung cancer (NSCLC) and to explore its prognostic value. Methods The expression of CCR9 was detected by immunohistochemistry in tumor tissues and corre-sponding adjacent normal tissues from 119 NSCLC patients. Additionally, the correlation between CCR9 expression and the clinicopath-ologic features of NSCLC and the relationship between prognostic factors and overall survival rate were analyzed by statistical methods. Results The positive expression rate of CCR9 was significantly higher in NSCLC tissues (54. 6%) than that in adjacent normal lung tissues (10. 1%) (P<0. 05). The expression of CCR9 in NSCLC was correlated with histopathologic type, lymph node status and p-TNM stage (P<0. 05). Kaplan-Meier survival analysis suggested that the positive expression of CCR9 was negatively correlated with the overall survival rate (Log-rank=9. 917, P=0. 002). Univariate analysis showed that the lymph node status, p-TNM stage and the positive expression of CCR9 made great difference to postoperative overall survival (P<0. 05). Multivariate analysis showed that CCR9 expression was an independent prognostic factor for overall survival of NSCLC patients ( RR=0. 447, 95%CI:0. 201 ~0. 993, P<0. 05). Conclusion The expression of CCR9 may predict a poor prognosis in the patients with NSCLC, so it can be used as a novel NSCLC biomarker.