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Contribution of PLCε/NF-κB Signal Pathway on Proliferation of Tumor Cells in Renal Cell Carcinoma

Author: WangChunYuan
Tutor: LuoChunLi
School: Chongqing Medical University
Course: Clinical Laboratory Science
Keywords: PLCε Renal cell carcinoma RT-PCR Proliferation 786-0 cells BAY11-7082 COX-2 c-myc
CLC: R737.11
Type: Master's thesis
Year: 2011
Downloads: 35
Quote: 0
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Abstract


The first part PLCε expression in RCC and its clinical significance Objective To investigate PLCε (phospholipase C epsilon) expression and clinicopathological relationship between renal cell carcinoma (Renal cell carcinoma, RCC). Methods RT-PCR assay the PLCε genes in renal cell carcinoma (n = 33) tissue and corresponding paraneoplastic tissue mRNA expression and analyze its relationship with clinical and pathological staging classification. Results PLCεmRNA expression levels in renal cell carcinoma was significantly higher than that of adjacent tissues (P lt; 0.001) PLCεmRNA expression with RCC clinical stage of about 0.05 (P lt; 0.05), with the histological grade (P gt; ). The conclusion PLCεmRNA significantly higher expression in renal cell carcinoma, and clinical stage, the speculated PLCε may be related to the development of RCC occurs. Second part PLCε the purpose of detection of renal cell carcinoma 786-0 the shRNA PLCε transfected cells of renal cell carcinoma 786-0 cell proliferation, cell proliferation and to analyze the possible molecular mechanisms by RT-PCR and Western The blot was renal cell carcinoma 786-0 cells transfected with shRNA PLCε PLCε of NF-κB, COX-2 and c-myc mRNA and protein expression changes. And Western detection of NF-κB nuclear protein and cytoplasmic protein expression changes. MTT and colony formation the assay shRNA PLCε 786-0 cell proliferation inhibition. The immunofluorescence detection the shRNA PLCε transfected 786-0 cells translocation of NF-κB nucleoprotein situation. The results of RT-PCR and Western blot results showed The the shRNA PLCε significantly inhibit the PLCε of NF-κB, COX-2 and c-myc mRNA and protein expression, and inhibits the cytoplasm and nucleus of NF-κB protein expression. MTT and colony formation assay results display The shRNA PLCε could significantly inhibit the proliferation of 786-0 cells. The immunofluorescence inhibit the of NF-ΚB protein expression, shRNA PLCε. Conclusion the shRNA PLCε may by down-regulating NF-kappa B, COX-2 and c-myc gene and protein expression to inhibit the proliferation of 786-0 cells. The third part of the NF-κB-specific inhibitor BAY11-7082 786-0 cell proliferation Objective To investigate the NF-κB specific inhibitor BAY11-7082 inhibit the proliferation of 786-0 cells and NF-κB protein, cytoplasmic and extracellular nucleoprotein as well as the expression of COX-2 and c-myc protein, is further validation PLCε to inhibit the proliferation of 786-0 cells via NF-κB and COX-2 and c-myc protein expression. Methods handle 786-0 cells with different concentrations of drugs BAY11-7082 MTT method and colony formation assay BAY11-7082 on cell proliferation, BAY11-7082 treated NF-κB protein were detected by Western blot, the nucleoprotein and cytoplasmic protein, COX-2 and c-myc protein expression. Immunofluorescence detection BAY11-7082 on NF-κB nuclear protein translocation. Results BAY11-7082 role inhibit 786-0 cell proliferation. Western blot analysis showed BAY11-7082 significantly inhibited the expression of COX-2 and c-myc protein (P lt; 0.01), but did not inhibit NF-κB protein expression (P gt; 0.05), therefore, we further detect NF -κB cytoplasm and nucleus of NF-κB protein found to suppress the expression of NF-κB nuclear proteins (P lt; 0.01). Conclusion BAY11-7082 may be through the inhibition of the nuclei NF-κB protein expression to inhibit COX-2 and c-myc gene and protein expression and thus achieve inhibition of proliferation of 786-0 cells.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Urinary tumors > Kidney,renal pelvis tumor
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