摘要
目的观察7次跨膜超蛋白家族成员4（TMTSF4）基因表达水平对乳腺癌细胞生物学行为的影响。方法构建TM7SF4慢病毒表达载体，转染MCF-7人乳腺癌细胞株，下调TM7SF4基因的表达，运用Cellomics、流式细胞术、碘化丙锭．荧光活化细胞分类计（PI-FACS）法及克隆形成实验，研究TM7SF4基因对NCF-7细胞的增殖、凋亡、细胞周期及克隆形成能力的影响。结果（1）对MCF-7细胞增殖能力的影响：MCF-7细胞连续培养5d后，实验组细胞数为：（239．1±37．7）、（508．4±52．8）、（282．2±84．0）、（308．0±167．1）、（131．6±82．6）个/孔；而空白细胞组细胞数为：（382．8±38．0）、（1115．0±155．9）、（1426．2±192．4）、（2468．4±232．6）、（2903．0±165．7）个/孔，空质粒组细胞数为：（380．8±39．0）、（1090．0±145．9）、（1426．0±180．3）、（2379．0±228．6）、（2855．0±156．6）个／孔，结果显示实验组细胞生长及细胞增殖倍数明显受到抑制，与空白对照组及空质粒组比较差异有统计学意义（P〈0．05）；（2）对细胞周期的影响：实验组MCF-7细胞处于G0／G1期、s期、G2／M期的细胞数为（61．87±0．27）、（20．31±0．11）、（17．83±0．35）个／孔；而空白对照组处于C0／G1期、S期、G2／M期的细胞数为（57．46±1．40）、（30．42±0．36）、（12．83±0．55）个／孔；空质粒组处于C0／G1期、S期、G2／M期的细胞数为（57．78±1．23）、（30．11±0．47）、（13．30±0．79）个／孔。实验组与空白对照组、实验组与空质粒组比较差异有统计学意义（P〈0．05）；（3）对细胞凋亡的影响：实验组散点图MCF-7细胞的凋亡率为（9．94±0．57）％，峰型图MCF-7细胞的凋亡率为（27．37±1．11）％；空白对照组散点图MCF-7细胞的凋亡率为（5．88±0．23）％，峰型图MCF-7细胞的凋亡率为（23．34±0．50）％；空质粒组散点图MCF-7细胞的凋亡率为（5．80±0．24）％，峰型图的凋亡率为（23．37±0．46）％，实验组与空白细胞对照组及实验组与空质粒组比较差异有统计学意义（P〈0．05）；（4）对细胞克隆形成的影响：实验组形成细胞克隆集落数目为（11±3）个，空白细胞对照组为（34士2）个，空质粒组为（29±1）个。实验组与空白细胞组及空质粒组比较差异有统计学意义（P〈0．05）。结论TM7SF4基因具有促进MCF-7乳腺癌细胞增殖、克隆形成的能力，同时抑制细胞凋亡及细胞周期进展。TM7SF4基因通过影响乳腺癌细胞生物学行为发挥促癌发生发展的作用。
Objective To investigate the effect of transmembrane 7 superfamily member 4 （TM7SF4） gene on behavior of breast cancer cell. Methods The TM7SF4 lentivirusis constructed and used to transfect MCF- 7 human breast cancer ceils, and then down- regulate TMTSF4 gene. Investigate the effect of TM7SF4 gene on the proliferation, apoptosis, cell cycle and cell proliferation of MCF -7 cells by Cellomics, flow cytometry, propidium iodide -fluorescence activated cell sorter （PI -FACS） and clone formation. Results （ 1 ） Effect on proliferation of MCF -7 cells： the number of MCF- 7 cellsof experi- mental group in 5 days cultured were ： （239. 1 ± 37.7 ）, （ 508.4 ± 52. 8 ）, （ 282. 2 ± 84. 0 ）, （ 308.0 ± 167. 1 ）, （ 131.6 ± 82. 6） cell/well; the number of MCF - 7 cells of no - treatment control group in 5 days cultured were： （382.8 ± 38.0）, （1 115.0± 155.9）, （1426.2± 192.4）, （2468.4±232.6）,（ 2 903.0 ± 165.7 ） cell/well ; the number of MCF - 7 cells of plasmid - treatment group in 5 days cultured were： （380.8±39.0）, （1090.0±145.9）, （1 426.0±-180.3）, （2379.0±228.6）, （2855.0± 156. 6） cell/well, results showed that the number of cells and rate of cells growth of experimental group were significantly inhibited （P 〈0. 05） ; （2） Effect on cell cycle of MCF-7 cells： the number of MCF -7 cells of experimental group at G0/G± stage, S stage, GJM stage were （ 61.87 ± 0. 27 ）, （20. 31 ±- 0. 11 ） , （17. 83 ±0. 35） cell/well, respectively; and that of no - treatment control group were （57.46 ± 1.40）, （30.42 s-0. 36）, （ 12. 83 ± 0. 55） cell/well, respectively, and that of plasmid - treatment group were （57. 78 ± 1.23）, （30. 11 ±0. 47）, （ 13.30 ±0. 79） cell/well, respectively. Statistical differences were found between experimental group and no -treatment control group, between experimental group and plus- mid -treatment group （P 〈0. 05） ; （3） Effect on apoptosis of MCF -7 cells： MCF -7 cell apoptosis rate of experimental group, no - treatment control group and plasmid - treatment group was ： （ 9.94 ± 0. 57 ） %, （5.88±0.23）%, （5.80 ±0.24）% in scatter diagram and （27.37±1.11）%, （23.34±0.50）%, （23.37± 0. 46）% in peak valuesdiagram, respectively; There were statistical difference between experi- mental and no - treatment control group, between experimental group and plasmid - treatment group both in scatter diagram （ P 〈 0.05 ） and peak values diagram （ P 〈 0. 05 ） , respectively ; （4） Effect on clone for- mation of MCF - 7 ceils ： number of the colony MCF - 7 cells in the experimental group, no - treatment control group and plasmid - treatment control was 11 ± 3, 34 ± 2, 29 ± 1, respectively. There were statisti- cal difference between experimental group and no - treatment control group （ P 〈 0.05 ） , between experi- mental group and plasmid - treatment control group （ P 〈 0. 05 ） , respectively. Conclusion It was con- firmed that the TM7SF4 gene had the ability to promote the proliferation and clone formation of MCF - 7 breast cancer cells, and to inhibit the apoptosis and cell cycle progression. TM7SF4 gene played a role in promoting the development of breast cancer cells. A new theoretical basis and experimental data were pres- ented for early diagnosis and targeted therapy of breast cancer.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第12期2691-2694,共4页
Chinese Journal of Experimental Surgery
基金
河南省医学科技计划项目（201503117）
关键词
乳腺癌
7次跨膜超蛋白家族成员4
凋亡
增殖
Breast cancer
Transmembrane 7 superfamily member 4
Apoptosis
Prolifera- tion
作者简介
通信作者：张威，Email：xianxu77@163．com
