Adenovirus-mediated Mda-7/il-24 Gene On Apoptosis Of Human Nasopharyngeal Carcinoma Cell Cne

MDA-7(melanoma differentiation-associated gene-7), also known as IL-24(Interleukin-24), was identified and cloned by Jiang using the differentiation induction subtraction hybridization approach after treatment of the human HO-1 melanoma cell line with IFN-βand MEZ resulted in growth arrest and terminal differentiation. Later it was re-designated as Interleukin-24(IL-24) by its gene structure and function of immune molecules. Nowadays it has been demonstrated that supraphysiological level's MDA-7 can induce growth arrest and apoptosis in different kinds of transformed cells, but has no similar effect on normal cells, and also the effect of immune stimulation, antiangiogenesis and improving the sensitivity of Radiotherapy and chemotherapy, etc. Significant experimental results are achieved from both in vivo and in vitro experiments and it has broad prospects for cancer therapy.Although nasopharyngeal carcinoma patients can be seen in many countries and regions among the five continents, the disease incidence is very small in most parts of the world which is usually below 1/100,000 and it has Regional and ethnic differences, since it's seen more in Asian than in Caucasian people. So although there have been much research on breast cancer, liver cancer, rectal cancer, etc, nasopharyngeal carcinoma as a common cancer in China especially the south of China, hasn't been researched on the apoptosis effect of MDA-7. Moreover there is worthy of attention that MDA-7 gene can not induce apoptosis in all of the transformed cells, some transformed cells such as pancreatic cancer cells and some of the lung cancer cells are resistant to the apoptosis effect of MDA-7.Adenovirus vector is now of a wide range of applications, though it is more complicated to construct compared to other virus vectors, Since the adenovirus vector system has the security, stability, and efficiency of cell infection and gene expression. We can achieve gene transferring to the target cells and tissues and correlative protein's expression and acting by cloning the target gene onto the recombinant adenovirus genome.In this paper we use a recombinant adenovirus 5(Ad5) vector (Replication -deficient) to construct recombinant virus Ad-mda7 and empty virus Ad-GFP, using HEK 293T cells for the virus production and amplification. The titer-measured virus are used to research on the apoptosis effect of nasopharyngeal carcinoma quantitatively. The result of Hoechst 33258 staining for apoptosis and FACS analysis (PI staining) suggested that: compared to Ad-GFP and PBS treating group, Ad-mda7 has evident apoptosis-inducing effect in the CNE cell, not only for the apoptotic morphology results but also for the accumulated proportion of SubG1 phase and G2/M-block phase in cell cycle FACS results, which all provided evidences for the future research and clinical applications.