Dissertation > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Bone marrow tumor

Comparison of Tranfection Efficiency Between Adenovirus Complexes and Liposome Complexes in Multiple Myeloma Cell Line

Author MaFangYu
Tutor LiuJing
School Central South University
Course Clinical
Keywords Multiple myeloma PDCD adenovirus Liposome transfection efficiency
CLC R733.3
Type Master's thesis
Year 2013
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Objective:Compare transfection efficiency of adenovirus rAd-PDCD5-EGFP complexes and pYr-ads-4-PDCD5complexes in multiple myeloma (MM) KM3cells, U266cells, Human chronic myelogenous leukemia K562cells, Hepatocellular carcinoma HepG2cells. Explore an optimum administration route for introducing exogenous PDCD5gene, which was regarded as a MM apoptosis-related gene, into MM cells.Methods:Eukaryotic expression plasmid pYr-ads-4-PDCD5complexes and recombinated adenovirus rAd-PDCD5-EGFP complexes were built. Then transfect these two complexes into KM3cells, U266cells, K562cells, HepG2cells respectively. Three groups were settled, control group (not transfected/infected cells Con), negtive control group (transfected vector cells or infected negative control adenovirus cells NC), PDCD5group (transfected PDCD5recombinated vector cells or infected PDCD5recombinated adenovirus cells).48h latter, transfection/infection efficiency was observed by fluorescence microscope. Meanwhile, PDCD5mRNA level were detected by Real-time Quantitative PCR and PDCD5protein expression level was measured by Western blot among those four types of cells seperately.Result:The transfection efficiency of Liposome mediated PDCD5recombinant plasmid in KM3, U266and K562were45%,49%,55%respectively, while about70%in HepG2cells. Compare to control and NC group, both PDCD5mRNA and protein expression level were significantly increased in PDCD5group. Additionally, the adenovirus rAd-PDCD5-EGFP mediated transfection efficiency in KM3and U266cells was33%,29%, while in K562cells was65%. Surprisingly, even higher infection efficiency (about80%)was detected in HepG2compare to other three types of cells. The results of Real-time PCR and Western blot showed that PDCD5mRNA and protein level in PDCD5group were significantly increased compare to control and NC group, while there were no significant difference between control group and NC group.Conclusions:Based on our experimental results, we conclude that the Liposome-mediated transfection efficiency was higher than adenovirus-mediated infection efficacy in MM KM3, U266cells. Consequently, liposome may be an ideal vector for introducing exogenous gene into MM cells, and it provide a powerful tool or theoretical basis for further study PDCD5gene-induced apoptosis mechanisms of MM.

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