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Inhibitory Effects of Cyclooxygenase-2 Inhibited by shRNA on the Growth and Angiogenesis of Human Liver Cancer Cell Subcutaneous Xenograft Tumors in Nude Mice

Author: ChenGuangHui
Tutor: TuZhiGuang；LiuYu
School: Chongqing Medical University
Course: Clinical Laboratory Science
Keywords: Cyclooxygenase-2 Restructuring interference plasmid pshRNA - Cox - 2 Subcutaneous tumor Angiogenesis Apoptosis
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 27
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Abstract


Objective: To investigate restructuring interference the plasmid pshRNA- Cox - 2 in human hepatoma cells Hep3B nude mice tumor growth and tumor angiogenesis inhibitory effect on tumor cell apoptosis role in promoting . METHODS: The recombinant interference plasmid pshRNA - COX - 2 and negative control plasmid pshRNA - HK identified by sequencing were transfected Hep3B cells , G418 - resistant screening monoclonal cell lines , RT - PCR and Western blot were used to detect COX-2 mRNA RT-PCR, expression of VEGF mRNA and protein expression . The the Hep3B monoclonal cell lines and non-transfected Hep3B cells were grown in nude mice , measured every 3 days once tumor size, tumor growth curve . After 4 weeks , the mice were sacrificed and tumor volume was measured by RT - PCR and Western blot were used to detect tumor organization COX - 2 mRNA and protein expression , immunohistochemical analysis of tumor tissue COX-2 protein expression and microvessel density ( MVD ) electron microscopy and TUNEL assay of tumor cell apoptosis . Results: Compared with the non-transfected cells , the interference group COX-2 mRNA and protein expression inhibition rates were 65.3% and 52.8 % ( P lt ; 0.05 ) , knockdown expression of VEGF mRNA inhibition rate was 56.5 % ( P lt ; 0.05 ). Plasmid compared to cells transfected with negative control knockdown COX-2 mRNA and protein expression inhibition rates were 54.6% and 43.5 % ( P lt ; 0.05 ) , the interference group VEGF mRNA expression inhibition rate was 44.6 % ( P lt ; 0.05). Interference group final tumor size was significantly less than the negative group and blank group ( P lt ; 0.01 ) . Compared with the control group , the interference tumor tissue in COX-2 mRNA and protein expression inhibition rates were 48.5% and 61.4 % ( P lt ; 0.05 ) . Compared with the negative group , interference tumor tissue in COX-2 mRNA and protein expression inhibition rates were 41.8% and 55.7 % ( P lt ; 0.05 ) . Interference group COX-2 score and MVD were significantly lower than that of the negative and blank control group ( P lt ; 0.01 ) , electron microscopy showed interference apoptotic cells increased TUNEL method knockdown tumor cell apoptosis index was significantly higher than that in the negative group and blank group ( P lt ; 0.01 ) . Conclusion: Restructuring interference plasmid pshRNA - COX - 2 inhibition by targeting COX-2 expression was significantly inhibited human hepatoma Hep3B cells transplanted into nude mice and tumor growth and tumor angiogenesis , and while significantly promote tumor cell apoptosis .

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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