Effect Of MCL-1 Antisense Oligonucleotide On The Reversion Of Drug Resistance Of Malignant Melanoma Cells.

Objective Malignant melanoma is the most aggressive form of skin cancer. Multidrug resistance is the vital factor of ineffective treatment in malignant melanoma. Chemotherapeutic agents function mainly by inducing apoptosis in tumor cells. While over expression of the antiapoptotic gene can inhibit the effect of the chemotherapy on melanoma cells, cause the poor prognosis and high death rate of melanoma. The B-cell lymphoma-leukemia 2(Bcl-2) protein family represents one of the major groups of apoptosis regulatory proteins, sharing the same structural characteristics. At least 15 Bcl-2 family members have been identified in mammalian cells . Despite their structural similarities, Bcl-2 family members can either facilitate cell survival or promote cell death. MCL-1 is a 37-kDa protein originally MCL-1(myeloid cell leukemia-1) belongs to the pro-survival Bcl-2 subfamily. Antisense technique is the highlight with its effectiveness, specificity and convenience. This experiment transfected MCL-1 antisense oligonucleotide(AS-ODN) into WM451 cells with lipofectamine 2000 to decrease the expression of MCL-1, at the same time used chemotherapy in combination, to observe the effect of MCL-1 AS-ODN on the chemoresistance of melanoma.Methods MCL-1 AS-ODN was transfected into cultured melanoma cell line WM451 with lipofectamine 2000. The expression of Mcl-1 nucleotide and protein was detected by Reverse transcription polymerase chain reaction (RT-PCR),Western blot and Immunocytochemistry, and apoptosis was assayed by Electron microscope and FAC. The sensitivity of WM451 cells to chemotherapeutic agents was assayed by MTT method.Results After 2 hours transfection, the expression of mRNA...