PDGF-α受体沉默对人晶状体上皮细胞增殖和凋亡的影响
发布时间：2018-12-15 04:28
【摘要】：目的研究血小板源性生长因子-α受体沉默对人晶状体上皮细胞增殖及凋亡的影响,为治疗后发性白内障提供实验依据。 方法1、体外培养人晶状体上皮细胞株SRA01/04,使用阳离子脂质体将合成的血小板源性生长因子-α受体反义寡核苷酸(PDGFR-α ASODN)转染入细胞株SRA01/04,倒置显微镜下观察SRA01/04的形态学改变。2、按照实验分组(对照组、错义寡核苷酸组、反义寡核苷酸组)PDGFR-α ASODN作用于SRA01/04细胞24h、48h、72h后,MTT法检测细胞的增殖。3、根据实验分组将PDGFR-α ASODN作用于SRA01/04细胞48h后,流式细胞仪检测PDGFR-α ASODN对SRA01/04细胞周期及细胞凋亡的影响；Hoechst33258荧光染料染色法分析细胞凋亡；4、RT-PCR检测PDGFR-α ASODN作用48h后PDGF-α受体表达的变化情况。 结果1、PDGFR-α ASODN转染SRA01/04细胞48h后,实验组大部分细胞皱缩、变圆、甚至碎裂,死亡细胞漂浮于培养液中,部分活细胞分裂相变少、贴壁不牢、胞体边界模糊,有的甚至失去了原来的形状；与低浓度药物组比较,高浓度药物组对细胞的抑制作用增强。2、PDGFR-α ASODN作用于SRA01/04细胞24h～72h, SRA01/04细胞增殖受抑制,且随时间的延长抑制作用增强,实验组较对照组和错义寡核苷酸组比较对细胞的抑制作用增强,差异有统计学意义(P㩳0.05)。3、PDGFR-α ASODN能诱导SA.R01/04细胞凋亡,PDGFR-α ASODN作用于SRA01/04细胞48h后实验组细胞凋亡率分别为(3.22±0.25)%、(5.29±0.27)%,与对照组(0.75±0.67)%和错义寡核苷酸组(1.46±0.60)%比较,差异有统计学意义(P0.05)；实验组G1期细胞分布率分别为(53.31±1.30)%、(59.98±0.95)%,与对照组(47.73±1.18)%和错义寡核苷酸组(49.48±1.09)%比较,差异有统计学意义(P0.05)。4、PDGFR-α ASODN作用于SRA01/04细胞48h后,Hochest33258染色结果表明：实验组细胞核固缩、核浓染、致密凋亡细胞与对照组比较增多。5、PDGFR-α ASODN作用于SRA01/04细胞48h后,PDGF-α受体在SRA01/04中表达下调。 结论1、PDGF-α受体反义寡核苷酸能抑制人晶状体上皮细胞的增殖,诱导其凋亡。2、PDGF-α受体反义寡核苷酸沉默SRA01/04中PDGF-α受体基因的表达。3、PDGF-α受体在人晶状体上皮细胞增殖过程中起重要作用。
[Abstract]:Objective to study the effect of platelet-derived growth factor-伪 receptor silencing on the proliferation and apoptosis of human lens epithelial cells. Methods 1. Human lens epithelial cell line SRA01/04, was transfected into SRA01/04, cell line by using cationic liposome to transfect the synthesized antisense oligodeoxynucleotide of platelet-derived growth factor- 伪 receptor (PDGFR- 伪 ASODN) into the cell line SRA01/04, in vitro. The morphologic changes of SRA01/04 were observed under inverted microscope. (2) PDGFR- 伪 ASODN (control group, missense oligonucleotide group, antisense oligonucleotide group) was treated with PDGFR- 伪 ASODN for 24 h, 48 h and 72 h, respectively. MTT assay was used to detect the proliferation of SRA01/04 cells. 3. After treated with PDGFR- 伪 ASODN for 48 h, the effects of PDGFR- 伪 ASODN on SRA01/04 cell cycle and apoptosis were detected by flow cytometry. Hoechst33258 fluorescent dye staining was used to detect apoptosis, and RT-PCR was used to detect the expression of PDGF- 伪 receptor after 48 h of PDGFR- 伪 ASODN treatment. Results 1After transfection of PDGFR- 伪 ASODN into SRA01/04 cells for 48 h, most of the cells in the experimental group were shrinked, rounded or even broken, the dead cells were floating in the culture medium, some of the living cells had less mitotic phase change, the adherent was not firm, and the boundaries of the cells were blurred. Some even lost their original shape; Compared with the low concentration drug group, the inhibitory effect of high concentration drug group on SRA01/04 cells was enhanced. 2 PDGFR- 伪 ASODN inhibited the proliferation of SRA01/04 cells for 24 h and 72 h, and the inhibitory effect was enhanced with the prolongation of time. Compared with the control group and the missense oligonucleotide group, the inhibitory effect of the experimental group on SA.R01/04 cells was significantly increased (P0. 05). 3 PDGFR- 伪 ASODN could induce the apoptosis of SA.R01/04 cells. After treated with PDGFR- 伪 ASODN for 48 h, the apoptotic rates of SRA01/04 cells in the experimental group were (3.22 卤0.25)% and (5.29 卤0.27)%, respectively, compared with those in the control group (0.75 卤0.67)% and missense oligonucleotide group (1.46 卤0.60)%. The difference was statistically significant (P0.05). The distribution rates of G1 phase cells in the experimental group were (53.31 卤1.30)% and (59.98 卤0.95)%, respectively, compared with those in the control group (47.73 卤1.18)% and missense oligonucleotide group (49.48 卤1.09)%. The difference was statistically significant (P0.05). (4) after treated with PDGFR- 伪 ASODN for 48 h, the results of Hochest33258 staining showed that the nuclei of the experimental group were pyknosis, dense staining and dense apoptotic cells were more than those of the control group. After PDGFR- 伪 ASODN was treated with SRA01/04 cells for 48 h, the expression of PDGF- 伪 receptor was down-regulated in SRA01/04. Conclusion 1 the antisense oligonucleotide of PDGF- 伪 receptor can inhibit the proliferation of human lens epithelial cells and induce its apoptosis. 2 the antisense oligonucleotide of PDGF- 伪 receptor silenced the expression of PDGF- 伪 receptor gene in SRA01/04. PDGF- 伪 receptor plays an important role in the proliferation of human lens epithelial cells.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R776.1
[Abstract]:Objective to study the effect of platelet-derived growth factor-伪 receptor silencing on the proliferation and apoptosis of human lens epithelial cells. Methods 1. Human lens epithelial cell line SRA01/04, was transfected into SRA01/04, cell line by using cationic liposome to transfect the synthesized antisense oligodeoxynucleotide of platelet-derived growth factor- 伪 receptor (PDGFR- 伪 ASODN) into the cell line SRA01/04, in vitro. The morphologic changes of SRA01/04 were observed under inverted microscope. (2) PDGFR- 伪 ASODN (control group, missense oligonucleotide group, antisense oligonucleotide group) was treated with PDGFR- 伪 ASODN for 24 h, 48 h and 72 h, respectively. MTT assay was used to detect the proliferation of SRA01/04 cells. 3. After treated with PDGFR- 伪 ASODN for 48 h, the effects of PDGFR- 伪 ASODN on SRA01/04 cell cycle and apoptosis were detected by flow cytometry. Hoechst33258 fluorescent dye staining was used to detect apoptosis, and RT-PCR was used to detect the expression of PDGF- 伪 receptor after 48 h of PDGFR- 伪 ASODN treatment. Results 1After transfection of PDGFR- 伪 ASODN into SRA01/04 cells for 48 h, most of the cells in the experimental group were shrinked, rounded or even broken, the dead cells were floating in the culture medium, some of the living cells had less mitotic phase change, the adherent was not firm, and the boundaries of the cells were blurred. Some even lost their original shape; Compared with the low concentration drug group, the inhibitory effect of high concentration drug group on SRA01/04 cells was enhanced. 2 PDGFR- 伪 ASODN inhibited the proliferation of SRA01/04 cells for 24 h and 72 h, and the inhibitory effect was enhanced with the prolongation of time. Compared with the control group and the missense oligonucleotide group, the inhibitory effect of the experimental group on SA.R01/04 cells was significantly increased (P0. 05). 3 PDGFR- 伪 ASODN could induce the apoptosis of SA.R01/04 cells. After treated with PDGFR- 伪 ASODN for 48 h, the apoptotic rates of SRA01/04 cells in the experimental group were (3.22 卤0.25)% and (5.29 卤0.27)%, respectively, compared with those in the control group (0.75 卤0.67)% and missense oligonucleotide group (1.46 卤0.60)%. The difference was statistically significant (P0.05). The distribution rates of G1 phase cells in the experimental group were (53.31 卤1.30)% and (59.98 卤0.95)%, respectively, compared with those in the control group (47.73 卤1.18)% and missense oligonucleotide group (49.48 卤1.09)%. The difference was statistically significant (P0.05). (4) after treated with PDGFR- 伪 ASODN for 48 h, the results of Hochest33258 staining showed that the nuclei of the experimental group were pyknosis, dense staining and dense apoptotic cells were more than those of the control group. After PDGFR- 伪 ASODN was treated with SRA01/04 cells for 48 h, the expression of PDGF- 伪 receptor was down-regulated in SRA01/04. Conclusion 1 the antisense oligonucleotide of PDGF- 伪 receptor can inhibit the proliferation of human lens epithelial cells and induce its apoptosis. 2 the antisense oligonucleotide of PDGF- 伪 receptor silenced the expression of PDGF- 伪 receptor gene in SRA01/04. PDGF- 伪 receptor plays an important role in the proliferation of human lens epithelial cells.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R776.1
【参考文献】
相关期刊论文 前10条
1 宋艳萍,丁琴,刘琼,赵霞;Nd:YAG激光切除儿童后发性白内障50例[J];国际眼科杂志;2005年03期
2 陶津华;安小玲;孔郡;张瑞君;张劲松;;EGF体外对人晶状体上皮细胞迁移的影响[J];国际眼科杂志;2006年01期
3 王建明;孙乃学;冯海晓;熊蕾;惠娜;范雅稚;;Nd∶YAG激光两种切开方法治疗儿童后发性白内障的临床研究[J];国际眼科杂志;2008年08期
4 严浩;邱庆华;王艳丽;;EGFR反义寡核苷酸对人视网膜神经胶质细胞迁移的影响[J];国际眼科杂志;2010年08期
5 邱梅园;彭燕一;黄岚珍;张玉明;向秋;;PDGF-α受体反义寡核苷酸对体外视网膜色素上皮细胞增殖和凋亡的影响[J];国际眼科杂志;2011年02期
6 柯治生;蔡小军;邓小艳;;白细胞介素-1β反义寡核苷酸脂质体抑制后发性白内障的实验研究[J];武汉大学学报(医学版);2006年02期
7 倪琦;曾思恩;谭宁;郭芳;张美艳;;姜黄素对肝癌Hep1细胞增殖及凋亡的影响[J];山东医药;2010年07期
8 孔s，
本文编号：2379974
本文链接：https://www.wllwen.com/yixuelunwen/wuguanyixuelunwen/2379974.html
最近更新
教材专著
