Therapy Of Melanoma By Nodal-Cripto-1Signal Molecules

In recent years, the incidence of malignant melanoma is growing rapidly, thus ithas become the third largest type of skin cancer. Due to its high risk of prognosis andhigh rate of tumor metastasis, people can hardly cure this disease.During the past several decades, there has been a substantial increase in theincidence of cutaneous melanoma among all caucasian populations. The number ofdeaths due to cutaneous malignant melanoma has also increased in most fair-skinnedpopulations throughout the world in the past few decades.Recent studies show that embryonic microenvironment can reverse the malignantphenotype of tumor cells, and in this study, Nodal-Cripto-1signaling molecules play adecisive role. Based on the tissue organization field theory and the epigenetics theory,we establish the cell factor release system which will be able to secret Nodalinhibitor--Lefty and the monoclonal antibody of Nodal receptor--Cripto-1bymicroencapsulation technology. By in vivo and in vitro experiments, we successfullyconfirmed that the delivery system can inhibit the growth of B16tumor.First of all,we built the HEK293-pCDNA3.1-LEFTY1and HEK293-IRES-GFP-LEFTY2cell lines by cell transfection technology and confirmed that the celllines can stably express Lefty1and Lefty2.we also built the anti-Cripto-1-KVFELGLVAGLGHQE polypeptide monoclonal antibody cell lines(named2B11and2H7)by hybridoma technique,testing that their ELISA titer was1:32000and1:16000.Constructed using the high-voltage electrostatic device in our lab, we successfullyfabricated the gelatin-alginate microcapsules and the diameter can be controlled in200μm; To ensure the structural stability of microcapsules, We coated the microcapsuleswith poly-lysine and alginate according to the principle of positive and negative ionicbond. On the experiment, we proved that the microcapsules can exist in ICR mice threeweeks or more.Using this cell microcapsules by in vitro experiments, we found that: Lefty1canpromote apoptosis of B16cells and reversal B16tumor cells to low levels of malignantmelanoma-like cells, thereby inhibiting the tumor growth;2B11can inhibit B16tumorangiogenesis and reduce tumor cell invasion and metastasis, and thus significantlyinhibited B16tumor growth and exhibit cure trend.An alternative approach to treating cancer through systemic delivery of a tumor-targeted cytokine with microencapsulation of recombinant cells is a feasible goal.The many advantages of this delivery system are further enhanced by the platformnature of this technology, amenable to the delivery of molecules important in a varietyof pathways that can intervene cancer progression. For these reasons, delivery ofrecombinant gene products for cancer therapy via microencapsulation is a scientificallyand clinically viable concept.