PDSS2基因过表达影响结肠癌细胞生长的初步研究

发布时间：2018-11-12 20:36

【摘要】：目的:通过观察PDSS2基因过表达对人结肠癌SW480、HCT116细胞的细胞生长、细胞增殖、细胞周期、细胞凋亡、细胞迁移及侵袭能力等生物学行为的影响,为明确PDSS2与结肠癌的关系提供依据。方法:1、构建PDSS2过表达质粒(PC3.1-PDSS2);2、分别转染结肠癌SW480、HCT116细胞,实验分为两组:对照组(SW480组、HCT116组)转染空白质粒、实验组(PDSS2组)转染PDSS2过表达质粒;3、MTS检测细胞生长和增殖情况;4、流式细胞仪检测细胞周期、细胞凋亡率;5、Transwell法和划痕实验检测细胞迁移能力和细胞侵袭能力的变化。结果:(1)MTS检测细胞增殖:a、SW480实验组细胞d1、d2、d3增殖率分别为117.54%、161.19%、316.79%,其对照组为127.24%、206.72%、425.00%,差距具有统计学意义(P0.01);PDSS2过表达d1、d2、d3对SW4080细胞增殖的抑制率分别为7.62%、22.02%、25.46%,对照组为0.00%、0.00%、0.00%,差距具有统计学意义(P0.01)。b、HCT116实验组细胞d1、d2、d3增殖率分别为115.04%、150.81%、248.37%,其对照组为118.97%、164.43%、289.33%,差距具有统计学意义(P0.01);PDSS2过表达d1、d2、d3对HCT116细胞增殖的抑制率分别为5.98%、10.82%、16.53%,对照组为0.00%、0.00%、0.00%,差距具有统计学意义(P0.01)。(2)流式细胞仪检测细胞周期:a、SW480实验组G1期细胞占70.24%,其对照组59.54%;SW480实验组S期细胞25.00%、G2期4.77%,均低于其对照组的33.86%、6.60%;b、HCT116实验组G1期细胞占68.25%,其对照组57.10%;HCT116实验组S期细胞18.12%、G2期13.63%,均低于其对照组的24.14%、18.76%。(3)流式细胞仪检测细胞凋亡:a、SW480实验组细胞总凋亡率11.03%,其对照对照组为7.58%(P=0.012)。b、HCT116实验组细胞总凋亡率为16.03%,其对照组为13.01%(P=0.015)。(4)划痕实验显示培养48h后,a、SW480实验组细胞间距离为|445.62±27.45|,其对照组为|335.03±31.69|;b、HCT116实验组细胞间距离为|259.34±13.2|,其对照组为|188.38±9.5|。(5)Transwell法检测:a、SW480实验组迁移细胞数为136.5±13.25,其对照组为350.50±30.17,差距均具有统计学意义(P0.01);SW480实验组侵袭细胞数为59.17±9.85,其对照组为140.50±27.08,差距均具有统计学意义(P0.01);b、HCT116实验组迁移细胞数为19.83±2.32,其对照组为63.17±16.07,差距均具有统计学意义(P0.01);SW480实验组侵袭细胞数为8.83±1.72,其对照组为37.33±4.27,差距均具有统计学意义(P0.01)。结论:(1)PDSS2基因过表达可以抑制结肠癌细胞SW480、HCT116的增殖,阻止其从G1期向S期转化;(2)PDSS2基因过表达可以影响结肠癌细胞SW480、HCT116细胞周期的调节。(3)PDSS2基因过表达可以诱发结肠癌细胞SW480、HCT116的早期凋亡;(4)PDSS2基因过表达可以抑制结肠癌细胞SW480、HCT116的迁移和侵袭能力。
[Abstract]:Aim: to investigate the effects of overexpression of PDSS2 gene on cell growth, cell proliferation, cell cycle, apoptosis, migration and invasion of human colon cancer SW480,HCT116 cells. To provide the basis for clarifying the relationship between PDSS2 and colon cancer. Methods: 1Construction of PDSS2 overexpression plasmid (PC3.1-PDSS2), 2transfection of colon cancer SW480,HCT116 cells into two groups: control group (SW480 group, HCT116 group) transfected blank plasmid, experimental group (PDSS2 group) transfected PDSS2 overexpression plasmid; (3) MTS was used to detect cell growth and proliferation; (4) flow cytometry was used to detect cell cycle and apoptosis rate; (5) Transwell method and scratch test were used to detect the changes of cell migration and invasion ability. Results: (1) MTS was used to detect cell proliferation: the proliferative rate of the SW480 experimental group was 117.54 and 161.191.191.191.191.19. 79, respectively, and the control group was 127.240.206.72and 425.00%. The difference was statistically significant (P0.01). The inhibitory rate of PDSS2 overexpression on proliferation of SW4080 cells was 7.62 and 22.02and 25.46, respectively, and that of control group was 0.000.0.0.The difference was statistically significant (P0.01). B,). The proliferative rate of the HCT116 experimental group was 115.04 / 150.81 and 248.37, respectively, and the control group was 118.97 / 164.433.The difference was statistically significant (P0.01). The inhibitory rates of PDSS2 overexpression on the proliferation of HCT116 cells were 5.98 and 10.82 ~ 16.53, respectively, while those in the control group were 0.000.000. The difference was statistically significant (P0.01). (2) flow cytometry was used to detect the cell cycle: the G1 phase of ASW480 experimental group accounted for 70.24%, while that of the control group was 59.54; The cells in S phase of SW480 experimental group were 25.00 and 4.77 in G2 phase, which were lower than those in control group (33.86g / kg). The G1 phase of SW480 group was 68.25%, and that of control group was 57.10%. The S phase of HCT116 experimental group was lower than that of the control group (24.14 ~ 18.76). (3) flow cytometry was used to detect cell apoptosis: the total apoptosis rate of the experimental group was 11.03, and that of the control group was 11.03. The total apoptotic rate of the control group was 7.58% (P0. 012), the total apoptosis rate of the experimental group was 16. 03, and that of the control group was 13. 01% (P0. 015). (4) scratch test showed that after 48 hours of culture, a, the apoptosis rate was 16. 03% and 13. 01% (P0. 015). (4). The intercellular distance of SW480 group was 445.62 卤27.45? s, while that of control group was 335.03 卤31.69? The intercellular distance was 259.34 卤13.2in the experimental group and 188.38 卤9.5in the control group. (5) Transwell assay showed that the number of migration cells in the experimental group was 136.5 卤13.25, and that in the control group was 350.50 卤30.17. The differences were statistically significant (P0.01). The number of invasive cells in SW480 group was 59.17 卤9.85, while that in control group was 140.50 卤27.08. The difference was statistically significant (P0.01). The number of migration cells in BHCT116 experimental group was 19.83 卤2.32, while that in control group was 63.17 卤16.07.The difference was statistically significant (P0.01). The number of invasive cells in SW480 group was 8.83 卤1.72, while that in control group was 37.33 卤4.27. The difference was statistically significant (P0.01). Conclusion: (1) overexpression of PDSS2 gene can inhibit the proliferation of SW480,HCT116 and prevent its transformation from G1 phase to S phase. (2) overexpression of PDSS2 gene can affect the regulation of SW480,HCT116 cell cycle in colon cancer cells. (3) overexpression of PDSS2 gene can induce early apoptosis of SW480,HCT116 cells. (4) overexpression of PDSS2 gene can inhibit the migration and invasion of SW480,HCT116 in colon cancer cells.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.35

【相似文献】