The Construction Of Adenovirus Expressing TIPE2and The Effect Of TIPE2on The Proliferation Of Colon Cancer Cells

Objective The occurrence of colon cancer is closely associated with colonchronic inflammation.TIPE2(tumor necrosis factor-α-induced protein8-like2) is anewly identified negative regulator of inflammation and innate and adaptive immunity.This experiment aims to construct an adenovirus vector express ing TIPE2, and observethe effect of it on the proliferation of colon cancer cells.Methods Trizol method to extract the RNA in HEK293cells, reversetranscription, Primer star program amplification purpose fragment TIPE2-cDNA.Andthen TIPE2-cDNA was cloned into the transfer vector pShuttle-CMV,and recombinedwith adenoviral backbone vector pAdc68.The recombinant plasmids were packaged andamplified,purified in HEK293cells (named Adc68-TIPE2).The expression of TIPE2inHT29and SW620cells was examined by RT-PCR and Western blot. Proliferation ofcell was measured by MTT assay.Results TIPE2was successfully inserted into the adenoviral vector.After infectedwith Adc68-TIPE2, HT29and SW620cells expressed TIPE2higher than control. AfterHT29and SW620cells were treated by different concentration Adc68-TIPE2(108vp/ml,109vp/ml,1010vp/ml) for48hours,the proliferation rate of HT29cell was93%,71%, and14%, respectively.In HT29cell,the proliferation rate of groups treatedwith109vp/ml,1010vp/ml virus was significant lower than control(P<0.05),while thegroup treated with108vp/ml virus was not (P>0.05).The proliferation rate of SW620cell was89%,53%, and10%, respectively.In SW620cell,the proliferation rate of groupstreated with108vp/ml and109vp/ml virus was significant lower than control(P<0.05),while the group treated with1010vp/ml virus was not (P>0.05).Conclusion Adc68-TIPE2adenovirus vector was successfully constructed and itcan inhibit the proliferation of colon cancer HT29and SW620cells,which laid thefoundation for further studies of its antitumor mechanism.