Objective In this study, transfection efficiency of human papillomavirus (HPV) E7 DNA and protein constructs into HEK-293T normal cell line, and A549 and TC-1 tumor cell lines was evaluated by four delivery systems including supercharge GFP, hPP10 cell penetrating peptide, TurboFect and Lipofectamine using fluorescence microscopy and flow cytometry. Results The results indicated that Lipofectamine 2000 and TurboFect produced more effective transfection for GFP and E7-GFP DNA constructs in HEK-293T cells compared to in A549 and TC-1 cells (p < 0.05). In contrast, the supercharge GFP was efficient for E7 DNA and E7 protein delivery in both normal cell (similar to 83.94 and similar to 77.01% for HEK-293T), and cancer cells (similar to 71.69 and similar to 67.19% for TC-1, and similar to 73.86 and similar to 67.49% for A549), respectively. Indeed, in these cell lines, transfection efficiency by +36 GFP reached similar to 60-80%. Moreover, the hPP10 produced the best transfection result for E7-GFP protein in HEK-293T cells (similar to 63.66%) compared to TurboFect (similar to 32.95%); however, the efficiency level of hPP10 was only similar to 17.51 and similar to 16.36% in TC-1 and A549 cells. Conclusions Our data suggested that the supercharge GFP is the most suitable transfection vehicle for DNA and protein delivery into TC-1 and A549 tumor cell lines compared to other carriers.