Combined inhibitory effect of co-transfected siRNA plasmids in Hela cells on the expressions of glucose-6-phosphate dehydrogenase and transketolase like-1

Objective Glucose-6-phosphate dehydrogenase( G6PD) and transketolase like-1( TKTL1) are key regulatory enzymes of the pentose phosphate pathway( PPP) in tumors. This study observed the metabolic changes and biologic characteristics in tumor cells by silencing G6PD and TKTL1 genes,evaluated the combined inhibitory effect of co-transfected siRNA plasmids in Hela cells on G6PD and TKTL1 expressions,and investigated the effect and practicality of the combined suppression of PPP using co-transfected siRNA plasmids. Methods Specific siRNA expressing vectors targeting G6PD and TKTL1 genes were separately constructed and the recombinant plasmids were verified by restriction endonuclease and DNA sequencing. Then the recombinant plasmids were transfected into Hela cell line separately or jointly; their combined inhibitory effect on G6PD and TKTL1 expressions were evaluated by comparing their activities and mRNA expressions using RT-PCR. Results After joint transfection of the two types of recombinant plasmids,the G6PD and TKTL1 mRNA expressions were decreased significantly( 0. 96 ± 0. 05vs 0. 47 ± 0. 04,0. 98 ± 0. 05 vs 0. 41 ± 0. 04,both P 0. 01). Simultaneously,the activities of G6PD and TKT were also statistically decreased( 99. 2% vs 34. 5%,99. 8% vs 40. 1%,both P 0. 01).Compared with the single G6PD siRNA expression vector transfection group,G6PD mRNA expressions in the co-transfected group were remarkably reduced( 0. 47 ± 0. 04 vs 0. 31 ± 0. 04,P 0. 01). Conclusion Co-transfection of siRNA plasmids in Hela cells was an effective experimental program for the combined suppression of PPP.