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Fundamental Study on Treatment of Colon Carcinoma with Cellular Flice Inhibitory Protein Antisense Oligonucleotide Nanoparticles

Author: WenYouWu
Tutor: ZhuDaiHua
School: Chongqing Medical University
Course: Surgery
Keywords: cFLIP antisense oligonucleotide PAMAM-D colon carcinoma apoptosis
CLC: R735.35
Type: Master's thesis
Year: 2011
Downloads: 13
Quote: 0
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Abstract


Objective : To investigate the effect of cFLIP antisense oligoNucleotides(cFLIP-asODN) on the proliferation inhibition of human colon cancer cell line SW620;compare PAMAM-D with liposome LipofectamineTM2000 in cell transfection.Methods:PAMAM-D and cFLIP-asODN were mixed in different mass ratio to prepare carrier-antisense gene complex, and trasfected the human colon cancer cell line SW620. Meanwhile, liposome and cFLIP-asODN group, blank carrier and cFLIP-asODN group, liposome and nonsense oligonucleotides (nsODN) group and blank control group were set as comparison. After tranfection, fluorescence inverted microscope and flow cytometry(FCM) were used for transfection efficiency, Real-time PCR was used to observe the expression of cFLIP mRNA, MTT was used to test cell proliferation inhibition ratio of SW620, Western Blotting was used for the expression of cFLIP protein,and the flow cytometer for cellular apoptosis .Results:Comparing to the control groups, transfection efficiency of PAMAM-D-cFLIPasODN complex group was obviously increased(p<0.05), and the expressing of cFLIP mRNA was markedly decreased(p<0.05). After transfection, the cell proliferation was evidently inhibited (p<0.05) in complex group, the CPIR was higher than liposome cFLIP-asODN group(p<0.05).The expressing of cFLIP protein in PAMAM-D-cFLIPasODN complex group was apperantly decreased, that was lower than liposome and cFLIP-asODN group(p<0.05). At the same time, we has observed more remarkable apoptosis in the PAMAM-D+cFLIP-asODN complex group than the liposome and cFLIP-asODN group(p<0.05).Conclusion : PAMAM-D,as a carrier-system,could transfect cFLIP-asODN into human colon cancer cell line SW620, so that induce apoptosis in SW620. Compared with liposome, PAMAM-D was more efficient in transfection. PAMAM-D, as a high-performance carrier-system for gene and drug, could have a potential value in the targeting gene therapy of colon carcinoma.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Intestinal neoplasms > Colon tumor
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