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RNA Interference with survivin Gene on Human Adenoid Cystic Carcinoma Angiogenesis and Tumor Growth

Author: FuKai
Tutor: YangJun
School: Third Military Medical University
Course: Clinical Stomatology
Keywords: Survivin RNA Adenoid cystic carcinoma xenografts Angiogenesis Apoptosis Tumor growth
CLC: R73-3
Type: Master's thesis
Year: 2011
Downloads: 54
Quote: 0
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Abstract


Objective: to observe RNA interference of Survivin gene on the impact of adenoid cystic carcinoma xenograft tumor angiogenesis. Survivin gene adenoid cystic carcinoma transplanted tumor apoptosis observed RNA interference. 3 observed Survivin gene RNA interference adenoid cystic carcinoma tumor growth. Methods: 1, adenoid cystic carcinoma in nude mice bearing human: random 30, 4 to 6 weeks of age, body weight 16 to 20 g, Balb / c male nude mice were divided into six groups, each group of five. ACC-2 cells: tumor cell group (the subcutaneous inoculation ACC-2 cells), the tumor injection group (inoculated subcutaneously with ACC-2 cells, 15d underwent RNA interference the Survivin recombinant plasmid tumor Week tumor injection), plasmid interference (subcutaneous the inoculation transfected plasmid survivin gene RNA interference ACC-2 cells); ACC-M cells: tumor cell group (subcutaneous inoculation ACC-M cells), tumor injection group (ACC-M cells were inoculated subcutaneously, 15d underwent RNA interference Survivin recombinant plasmid peritumoral tumor injection), plasmid interference group (ACC-M cells inoculated subcutaneously with survivin gene RNA interference plasmid transfection). Foster care in the Third Military Medical University animal laboratory, all nude mice 25 days after spinal cord transection method put to death, under sterile conditions, peel the tumor to remove the envelope, part of the tumor tissue fixed in paraformaldehyde solution paraffin package, to be done buried sliced ??back. Part of the tumor tissue quickly set preserved in liquid nitrogen, and total RNA was extracted ready to be the line in tumor tissues. By measuring the size of adenoid cystic carcinoma xenograft tumor volume observed in each group adenoid cystic carcinoma tumor growth. By immunohistochemistry groups adenoid cystic carcinoma xenografts microvascular parameters. 4 groups adenoid cystic carcinoma xenograft tumor pro-angiogenic factor VEGF expression was detected by RT-PCR. 5 groups adenoid cystic carcinoma transplanted tumor cells by TUNEL assay apoptosis. To explore the RNA interference survivin gene on human adenoid cystic carcinoma xenograft tumor angiogenesis and tumor growth. Results: 1 tumor cell group tumor growth rapidly increased exponentially. The tumor injection group in 15d before tumor growth rapidly increased exponentially 15d underwent RNA interference the Survivin recombinant plasmid for the tumor peritumoral injection of tumor growth slowed down, non-multiple growth. Plasmid interference group tumor is slow-growing, non-multiple growth. With the tumor injection group and plasmid interference group with tumor cells for tumor volume (Figure 1, 2, Tables 1, 2), the results showed that the first two groups adenoid cystic cancer xenograft tumor volume is small, The difference was statistically meaningful (p lt; 0.05). Tumor injection group and plasmid interference group was not statistically significant (P gt; 0.05). Show: by RNA interference the Survivin gene can effectively inhibit tumor growth of adenoid cystic carcinoma xenograft tumor. 2, group adenoid cystic carcinoma xenografts immunohistochemical tissue sections, respectively, with the cavity microvascular density of the tumor injection group and plasmid interference group, microvascular density, average peritubular perimeter, average lumen area observed under the microscope tumor cell group (Table 3 and 4), the results show that the first two groups of various microvessel parameter values ??were lower, the difference was statistically meaningful (P lt; 0.05), tumor injection group and plasmid interference group was not statistically significant (P gt; 0.05). : Survivin gene RNA interference can inhibit adenoid cystic carcinoma xenograft tumor angiogenesis, thereby blocking the adenoid cystic carcinoma xenograft tumor occurrence and development of the means of transport of nutrients and oxygen required. 3, respectively, with the tumor injection group, plasmid interference group with tumor cell group VEGF electrophoresis with relative optical density values ??(Figure 5,6 Table 5,6), the results show the tumor injection group and plasmids interference of VEGF electrophoretic bands relative optical density ratio are low, and the difference was statistically meaningful (P lt; 0.05) plasmid interference group and tumor injection group showed no statistical significance (P gt; 0.05). The Survivin gene could down adenoid cystic cancer xenograft tumors in vivo to promote expression of the angiogenic factors VEGF: RNA interference to inhibit adenoid cystic carcinoma xenograft tumor angiogenesis. 4, observed under a confocal microscope the group adenoid cystic carcinoma xenograft tumor TUNEL tissue sections were transplanted tumor injection group and plasmid interference group average fluorescence value of tumor apoptosis in tumor cells compared with tumor cells (Figure 7,8 7,8), the results show that the average fluorescence values ??were higher in the first two sets of apoptotic tumor cells. The difference statistically meaningful (P lt; 0.05), the the tumor injection group and plasmid interference group showed no statistical significance (P gt; 0.05). Show that: by RNA interference the Survivin gene can promote apoptosis of adenoid cystic carcinoma transplanted tumor cells, to suppress the purpose of adenoid cystic cancer xenograft tumor growth. Conclusion: Survivin gene via RNA interference could down adenoid cystic carcinoma xenograft tumor pro-angiogenic factor VEGF expression, inhibition of adenoid cystic carcinoma xenograft tumor angiogenesis in vivo. Survivin gene RNA interference can promote apoptosis of adenoid cystic carcinoma transplanted tumor cells transplant in tumor growth inhibition of adenoid cystic carcinoma. Targeting Survivin gene RNA interference technology, pro-angiogenic factor VEGF expression by down adenoid cystic cancer xenograft in vivo inhibition of adenoid cystic carcinoma transplanted tumor angiogenesis, through the promotion of adenoid cystic cancer xenograft tumor cell apoptosis, inhibition of tumor growth and development of adenoid cystic carcinoma xenograft tumor.

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