Effect Of ShRNA Silencing NNMT Expression On Proliferation Of Human Colon Cancer Cell HT-29

Objective:To study the effect of NNMT on tumor cell proliferation and look for new and potential targets for cancer therapy.Methods:Lentiviral vectors targeting for NNMT were constructed by molecular clone techniques and successful infected human colon cancer cell HT-29. The expressions of NNMT mRNA and protein were identified by Real-Time PCR and western blot assay before and after NNMT silenced. The proliferation of HT-29cells was detected by MTT assay. The cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Changes in cell tumorigenicity ability were analyzed by plate colony formation assay and xenograft tumor formation test in nude mice.Results:The expressions of NNMT mRNA and protein in HT-29cells were remarkable suppressed by lentiviral vectors targeting for NNMT. Results showed that the cell growth curve turned to right, cell apoptosis rate increased and cell cycle was arrested at G0/G1phase, while the colony formation rate and tumorigecity ability in nude mice decreased remarkably after NNMT silenced.Conclusions:The use of shRNA-mediated RNAi technology can effectively silence NNMT expression in HT-29cells. NNMT downregulation could remarkably inhibite the proliferation of cancer cells, which suggests NNMT as a potential target for cancer therapy.