Experimental Research On Co-transfection Of WTP53and RB44Gene Into HXO-RB44Cell By Ultrasound Microbubble

Objectives:To investigate the cell growth inhibit effect of tumorsuppressor gene wild type P53and RB94after transfection using anultrasound-targeted microbuble destruction(UTMD) way. To primarilydiscuss the expression of MICA、MICB on the cell line of HXO-RB44.Methods:1. We constructed P53and Rb94co-expression vector,the P53expression vector and the Rb94expression vector.2. we measured the cell growth inhibition between the different transfectiongroups..3. We examined the expression of MICA and MICB on HXO-Rb44cells line.Results:1. The targeted gene was proved to be cloned into vector plasmid withthe sequence of Rb94,wtP53gene respectively. The expression of each genewas examined by RT-PCR method. 2.The transfection was conducted by microbubble with continuouswave by the parameter of sound intensity of0.5W/cm2， time of30s.Compared with the mock and single gene transfection group, tumorcell co-transfected with wtP53and RB94have the lowest cell growth ratenamely, a most significant cell proliferation inhibition, and a highest rate ofapoptosis as well as a strongest expression of bax protain.3. Immunohistochemistry showed that the expression of MICA andMICB was elevated on the HXO-RB44cell line,this was also proved by theresult of western blot.ConclusionThis study showed gene transfection by the way of UTMD canprovide the expression of exogenous gene of RB94and P53in single genetransfection or gene co-transfection groups.Among those groups the RB94and P53co-transfection group has a strongest cell growth inhibition effectin retinoblastoma cell line HXO-RB44.Meanwhile transfection of singleRB94gene can upregulate the expression of the NKG2D ligand--MICA,MICB.Nevertheless this study merely build some basicresource,and still need to be further investigate before applied in clinical.