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Trb3 gene shRNA vector and on the tongue squamous cell carcinoma cell proliferation and apoptosis

Author: WenHaoJie
Tutor: SuiJun
School: Kunming Medical College
Course: Otorhinolaryngology
Keywords: TSCC Trb3 gene shRNA Eukaryotic expression vector Proliferation Apoptosis
CLC: R739.86
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
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Abstract


Objective: This project intends to build a shRNA targeting human Trb3 eukaryotic expression vector and stable expression Trb3 shRNA tongue squamous cell lines , testing its target gene silencing effect was observed on the tongue squamous cell carcinoma cell proliferation and apoptosis Tca8113 impact , and to explore its mechanism . Methods: molecular cloning technology to build targeting human Trb3 shRNA vectors pSUPER-shTrb3; retroviral interference method to establish stable gene endogenous Trb3 tongue squamous cell carcinoma ; Western Blot, Real-timePCR detect the expression of target genes Trb3 Flow cytometry to detect cell proliferation and apoptosis , Western Blot detection of apoptosis related proteins . Results: The enzyme digestion and sequencing confirmed successfully constructed shRNA targeting human Trb3 gene eukaryotic expression vector , the use of retroviral expression method successfully establish a stable Trb3 shRNA tongue squamous cell carcinoma , Real-time PCR, Western Blot confirmed reorganization plasmid cells Trb3 mRNA and protein expression levels relative to empty vector cells was significantly decreased (P lt; 0.05). Flow cytometry confirmed that under the conditions of endoplasmic reticulum stress interfere with endogenous Trb3 TSCC can promote cell proliferation, inhibition of apoptosis , and apoptosis-related proteins cleaved-caspase 3,7, cleaved-PARP downregulated . Conclusion: endoplasmic reticulum stress state , Trb3 involved Tca8113 of tongue squamous cell apoptosis , the cell model is to Trb3 gene-based head and neck squamous cell carcinoma (head and neck squamous cell carcinoma, HNSCC) targeting treatment of experimental basis .

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CLC: > Medicine, health > Oncology > Oral cavity, maxillofacial tumors > Tongue cancer, sublingual tumor
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