【摘要】：目的:通過觀察PDSS2基因過表達對人結(jié)腸癌SW480、HCT116細胞的細胞生長、細胞增殖、細胞周期、細胞凋亡、細胞遷移及侵襲能力等生物學(xué)行為的影響,為明確PDSS2與結(jié)腸癌的關(guān)系提供依據(jù)。方法:1、構(gòu)建PDSS2過表達質(zhì)粒(PC3.1-PDSS2);2、分別轉(zhuǎn)染結(jié)腸癌SW480、HCT116細胞,實驗分為兩組:對照組(SW480組、HCT116組)轉(zhuǎn)染空白質(zhì)粒、實驗組(PDSS2組)轉(zhuǎn)染PDSS2過表達質(zhì)粒;3、MTS檢測細胞生長和增殖情況;4、流式細胞儀檢測細胞周期、細胞凋亡率;5、Transwell法和劃痕實驗檢測細胞遷移能力和細胞侵襲能力的變化。結(jié)果:(1)MTS檢測細胞增殖:a、SW480實驗組細胞d1、d2、d3增殖率分別為117.54%、161.19%、316.79%,其對照組為127.24%、206.72%、425.00%,差距具有統(tǒng)計學(xué)意義(P0.01);PDSS2過表達d1、d2、d3對SW4080細胞增殖的抑制率分別為7.62%、22.02%、25.46%,對照組為0.00%、0.00%、0.00%,差距具有統(tǒng)計學(xué)意義(P0.01)。b、HCT116實驗組細胞d1、d2、d3增殖率分別為115.04%、150.81%、248.37%,其對照組為118.97%、164.43%、289.33%,差距具有統(tǒng)計學(xué)意義(P0.01);PDSS2過表達d1、d2、d3對HCT116細胞增殖的抑制率分別為5.98%、10.82%、16.53%,對照組為0.00%、0.00%、0.00%,差距具有統(tǒng)計學(xué)意義(P0.01)。(2)流式細胞儀檢測細胞周期:a、SW480實驗組G1期細胞占70.24%,其對照組59.54%;SW480實驗組S期細胞25.00%、G2期4.77%,均低于其對照組的33.86%、6.60%;b、HCT116實驗組G1期細胞占68.25%,其對照組57.10%;HCT116實驗組S期細胞18.12%、G2期13.63%,均低于其對照組的24.14%、18.76%。(3)流式細胞儀檢測細胞凋亡:a、SW480實驗組細胞總凋亡率11.03%,其對照對照組為7.58%(P=0.012)。b、HCT116實驗組細胞總凋亡率為16.03%,其對照組為13.01%(P=0.015)。(4)劃痕實驗顯示培養(yǎng)48h后,a、SW480實驗組細胞間距離為|445.62±27.45|,其對照組為|335.03±31.69|;b、HCT116實驗組細胞間距離為|259.34±13.2|,其對照組為|188.38±9.5|。(5)Transwell法檢測:a、SW480實驗組遷移細胞數(shù)為136.5±13.25,其對照組為350.50±30.17,差距均具有統(tǒng)計學(xué)意義(P0.01);SW480實驗組侵襲細胞數(shù)為59.17±9.85,其對照組為140.50±27.08,差距均具有統(tǒng)計學(xué)意義(P0.01);b、HCT116實驗組遷移細胞數(shù)為19.83±2.32,其對照組為63.17±16.07,差距均具有統(tǒng)計學(xué)意義(P0.01);SW480實驗組侵襲細胞數(shù)為8.83±1.72,其對照組為37.33±4.27,差距均具有統(tǒng)計學(xué)意義(P0.01)。結(jié)論:(1)PDSS2基因過表達可以抑制結(jié)腸癌細胞SW480、HCT116的增殖,阻止其從G1期向S期轉(zhuǎn)化;(2)PDSS2基因過表達可以影響結(jié)腸癌細胞SW480、HCT116細胞周期的調(diào)節(jié)。(3)PDSS2基因過表達可以誘發(fā)結(jié)腸癌細胞SW480、HCT116的早期凋亡;(4)PDSS2基因過表達可以抑制結(jié)腸癌細胞SW480、HCT116的遷移和侵襲能力。
[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 鹵27.45? s, while that of control group was 335.03 鹵31.69? The intercellular distance was 259.34 鹵13.2in the experimental group and 188.38 鹵9.5in the control group. (5) Transwell assay showed that the number of migration cells in the experimental group was 136.5 鹵13.25, and that in the control group was 350.50 鹵30.17. The differences were statistically significant (P0.01). The number of invasive cells in SW480 group was 59.17 鹵9.85, while that in control group was 140.50 鹵27.08. The difference was statistically significant (P0.01). The number of migration cells in BHCT116 experimental group was 19.83 鹵2.32, while that in control group was 63.17 鹵16.07.The difference was statistically significant (P0.01). The number of invasive cells in SW480 group was 8.83 鹵1.72, while that in control group was 37.33 鹵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.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R735.35

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