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Silence ClC-3 chloride channel gene on nasopharyngeal carcinoma cell cycle

Author: YeDong
Tutor: ChenLiXin
School: Jinan University
Course: Physiology
Keywords: C1C-3 chloride channel Cell Cycle siRNA Nasopharyngeal carcinoma cells Cell volume Cyclin D1
CLC: R739.63
Type: Master's thesis
Year: 2011
Downloads: 51
Quote: 0
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Abstract


Objective: To study the ClC-3 siRNA silencing C1C-3 chloride channel gene on nasopharyngeal carcinoma cell cycle. Methods: Using siRNA technique to suppress nasopharyngeal carcinoma (CNE-2Z) cells ClC-3 gene expression, flow cytometry siRNA transfection efficiency, whole-cell patch clamp technique volume-activated chloride current, image analysis software Q500MC measured and calculated cell volume, immunofluorescence C1C-3 protein and cyclin Dl protein in intracellular distribution, Western blot detection of ClC-3 protein and cyclin D1 protein expression, cell cycle distribution by flow cytometry. Results: (1) ClC-3 siRNA transfected CNE-2Z cell transfection rate (63.8 ± 3.8)% (n = 3). Western blot results showed that compared with the control group, with 100nmol / L of ClC-3 siRNA transfected NPC cells, ClC-3 chloride channel protein expression was decreased (60.9 ± 4.0)% (n = 3, P lt; 0.05), while the disordered siRNA group, transfection reagent control group C1C-3 protein expression did not change significantly (n = 3, P gt; 0.05); (2) to 47% hypotonic solution for extracellular bath When the blank control group cell volume-activated chloride current average current density was (74.2 ± 6.5) pA / pF (80 mV), ClC-3 siRNA group cell volume-activated chloride current average current density was (13.7 ± 4.1) pA / pF, decrease compared with the blank control group (81.5 ± 4.7)% (n = 5, P lt; 0.05); (3) ClC-3 siRNA group cell regulatory volume decrease (regulatory volume decrease, RVD) capacity significantly weakened, 25min hypotonic the RVD was (10.5 ± 4.8)% (n = 16), and control group (42.6 ± 2.8)% (n = 20) compared to, RVD decreased 75.4% (P lt; 0.01); ( 4) Compared with the control group, ClC-3 siRNA group G0/G1 phase cells from (56.8 ± 2.8)% to (69.9 ± 3.0)%, S phase cells from (32.1 ± 1.7)% is reduced to (23.5 ± 1.5)% (n = 3, P lt; 0.05, while the negative control siRNA disorder, transfection reagent in the control group had no significant changes in cell cycle distribution (P gt; 0.05), indicates that silence chloride channel C1C-3 gene expression increased cell cycle arrest at G0/G1 phase; (5) immunofluorescence and Western blot analysis showed that, compared with the control group of cells, ClC-3 siRNA cells of cyclin Dl protein expression decreased (40.3 ± 2.0)% (n = 3, P lt; 0.05), while the control group and the transfection reagent disorderly siRNA group of cyclin D1 protein expression was not significantly change (n = 3, P gt; 0.05), indicates that ClC-3 siRNA inhibits cyclin D1 protein Conclusion: nasopharyngeal carcinoma CNE-2Z cells C1C-3 chloride channel protein. ClC-3 chloride channel protein involved in cell cycle regulation, the regulation of cells from the G0/G1 phase into S phase one of the important factors . C1C-3 chloride channel may influence the regulatory volume decrease and regulation of cyclin D1 protein expression and affect cell cycle progression.

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CLC: > Medicine, health > Oncology > Department of Otolaryngology tumor > Pharyngeal tumors
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