PTEN基因通过Akt-mTOR对乳腺癌细胞增殖与凋亡的影响
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摘要
目的:探讨PTEN基因通过Akt-mTOR对乳腺癌细胞增殖与凋亡的影响。方法:人乳腺癌MDA-MB-231细胞随机分为两组:pcDNA3.0组与pcDNA3.0-PTEN组,分别转染pcDNA3.0质粒、pcDNA3.0-PTEN质粒2μg,转染48 h收集细胞。采用CCK-8法检测细胞存活率,双染法检测细胞凋亡,Western-blot检测细胞蛋白表达。结果:pcDNA3.0-PTEN组的细胞存活率低于pcDNA3.0组,对比差异有统计学意义(P<0.05)。与pcDNA3.0组对比,pcDNA3.0-PTEN组的细胞凋亡率显著上升,对比差异有统计学意义(P<0.05)。pcDNA3.0-PTEN组的PTEN蛋白表达量高于pcDNA3.0组,Akt、mTOR蛋白表达量低于pcDNA3.0组,对比差异有统计学意义(P<0.05)。结论:PTEN基因过表达可通过抑制Akt-mTOR信号通路,提高乳腺癌细胞凋亡指数,降低细胞增殖活性,从而发挥抑癌作用。
Objective:To investigate the effect of PTEN gene on proliferation and apoptosis of breast cancer cells by Akt-mTOR signaling pathway.Methods:Human breast cancer MDA-MB-231 cells were randomly divided into two groups:pcDNA3.0 group,pcDNA3.0-PTEN group,respectively and were transfected with pcDNA3.0 plasmid,pcDNA3.0-PTEN plasmid of 2 μg.The cell viability was detected by CCK-8 method.Apoptosis was detected by double staining,and cell protein expression was detected by Western-blot.Results:The cell viability of the pcDNA3.0-PTEN group was lower than that of the pcDNA3.0 group,and the difference were statistically significant(P<0.05).Compared with the pcDNA3.0 group,the apoptosis rates of the pcDNA3.0-PTEN group were increased significantly,and the difference was statistically significant(P<0.05).The expression of PTEN protein in pcDNA3.0-PTEN group was higher than that in pcDNA3.0 group,and the expression of Akt and mTOR protein was lower than that in pcDNA3.0 group,and the difference was statistically significant(P<0.05).Conclusion:Over-expression of PTEN gene can inhibit the Akt-mTOR signaling pathway,increase the apoptosis index of breast cancer cells and decrease the cell proliferation activity,and thus play a tumor suppressing effect.
Objective:To investigate the effect of PTEN gene on proliferation and apoptosis of breast cancer cells by Akt-mTOR signaling pathway.Methods:Human breast cancer MDA-MB-231 cells were randomly divided into two groups:pcDNA3.0 group,pcDNA3.0-PTEN group,respectively and were transfected with pcDNA3.0 plasmid,pcDNA3.0-PTEN plasmid of 2 μg.The cell viability was detected by CCK-8 method.Apoptosis was detected by double staining,and cell protein expression was detected by Western-blot.Results:The cell viability of the pcDNA3.0-PTEN group was lower than that of the pcDNA3.0 group,and the difference were statistically significant(P<0.05).Compared with the pcDNA3.0 group,the apoptosis rates of the pcDNA3.0-PTEN group were increased significantly,and the difference was statistically significant(P<0.05).The expression of PTEN protein in pcDNA3.0-PTEN group was higher than that in pcDNA3.0 group,and the expression of Akt and mTOR protein was lower than that in pcDNA3.0 group,and the difference was statistically significant(P<0.05).Conclusion:Over-expression of PTEN gene can inhibit the Akt-mTOR signaling pathway,increase the apoptosis index of breast cancer cells and decrease the cell proliferation activity,and thus play a tumor suppressing effect.
引文
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[11] Sizemore GM,Balakrishnan S,Thies KA,et al.Stromal PTEN determines mammary epithelial response to radiotherapy[J].Nat Commun,2018,9(1):2783.
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[13] He L,Lin XF.Expression of miR-183 on breast cancer and effect of miR-183 on invasion and migration ability of human breast carcinoma MCF-7 cells by PTEN pathway[J].Chinese Journal of Cancer Prevention and Treatment,2017,24(7):451-457.[何力,林秀峰.miR-183靶向TBX3调控PTEN通路对乳腺癌生物学行为影响[J].中华肿瘤防治杂志,2017,24(7):451-457.]
[14] Barrett MT,Lenkiewicz E,Malasi S,et al.The association of genomic lesions and PD-1/PD-L1 expression in resected triple-negative breast cancers[J].Breast Cancer Res,2018,20(1):71.
[15] Zhou XL,Zhu CY,Zhang SG,et al.NDRG2 inhibits migration of breast cancer cells through upregulating PTEN expression[J].Cancer Research on Prevention and Treatment,2017,44(1):5-10.[周晓雷,朱重悦,张世光,等.NDRG2上调PTEN表达抑制乳腺癌细胞迁移的实验[J].肿瘤防治研究,2017,44(1):5-10.]
[16] Gu J,Wang Y,Wang X,et al.Downregulation of lncRNA GAS5 confers tamoxifen resistance by activating miR-222 in breast cancer[J].Cancer Lett,2018,30(434):1-10.
[2] Jin T.Research propress in signaling pathways and molecular mechanisms of brain metastases in breast cancer[J].Practical Oncology Journal,2017,31(5):468-471.[靳团.乳腺癌脑转移的信号通路和分子机制的研究进展[J].实用肿瘤学杂志,2017,31(5):468-471.]
[3] Li J,Gong X,Jiang R,et al.Fisetin inhibited growth and metastasis of triple-negative breast cancer by reversing epithelial-to-mesenchymal transition via PTEN/Akt/GSK3β signal pathway[J].Front Pharmacol,2018,31(9):772.
[4] Luo ZX,Chen YX,Ma ZY,et al.Expression of PTEN and VEGF-D in breast cancer[J].Journal of Modern Clinical Medicine,2017,43(2):131-132.[罗赵鑫,陈仰新,马智勇,等.乳腺癌组织中PTEN与VEGF-D的表达[J].现代临床医学,2017,43(2):131-132.]
[5] Plos One Editors.Retraction:Functional role of mTORC2 versus integrin-linked kinase in mediating Ser473-Akt phosphorylation in PTEN-negative prostate and breast cancer cell lines[J].PLoS One,2018,13(8):e0202299.
[6] Wan ZL,Liu C.Expression of PTEN protein and its relationship with prognosis in three patients with breast cancer[J].Shandong Medical Journal,2017,57(37):40-42.[万珍玲,刘超.三阴乳腺癌组织PTEN蛋白表达变化及与患者预后的关系[J].山东医药,2017,57(37):40-42.]
[7] Mollazadeh H,Afshari AR,Hosseinzadeh H.Review on the potential therapeutic roles of Nigella sativa in the treatment of patients with cancer:Involvement of apoptosis:Black cumin and cancer[J].J Pharmacopuncture,2017,20(3):158-172.
[8] Wang JJ,Fan ZR,Li LF,et al.Influence of HPK1 overexpression in proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells and its mechanism[J].Journal of Jilin University(Medicine Edition),2017,43(5):910-917.[王娇娇,范智蕊,李砺锋,等.HPK1过表达对乳腺癌MCF-7和MDA-MB-231细胞增殖和凋亡的影响及其机制[J].吉林大学学报(医学版),2017,43(5):910-917.]
[9] Han L,Zhang HC,Li L,et al.Downregulation of long noncoding RNA HOTAIR and EZH2 induces apoptosis and inhibits proliferation,invasion,and migration of human breast cancer cells[J].Cancer Biother Radiopharm,2018,33(6):241-251.
[10] Yang JM,Yu HJ,He Z.Expression and significance of PTEN and mTOR in HER2 overexpressing ductal carcinoma in situ[J].Guangdong Medical Journal,2017,38(14):2165-2168,2172.[杨剑敏,于海静,何舟.PTEN、mTOR在HER2过表达乳腺导管内癌中的表达及意义[J].广东医学,2017,38(14):2165-2168,2172.]
[11] Sizemore GM,Balakrishnan S,Thies KA,et al.Stromal PTEN determines mammary epithelial response to radiotherapy[J].Nat Commun,2018,9(1):2783.
[12] Wang L,Yang C,Liu XB,et al.B7-H4 overexpression contributes to poor prognosis and drug-resistance in triple-negative breast cancer[J].Cancer Cell Int,2018,13(18):100.
[13] He L,Lin XF.Expression of miR-183 on breast cancer and effect of miR-183 on invasion and migration ability of human breast carcinoma MCF-7 cells by PTEN pathway[J].Chinese Journal of Cancer Prevention and Treatment,2017,24(7):451-457.[何力,林秀峰.miR-183靶向TBX3调控PTEN通路对乳腺癌生物学行为影响[J].中华肿瘤防治杂志,2017,24(7):451-457.]
[14] Barrett MT,Lenkiewicz E,Malasi S,et al.The association of genomic lesions and PD-1/PD-L1 expression in resected triple-negative breast cancers[J].Breast Cancer Res,2018,20(1):71.
[15] Zhou XL,Zhu CY,Zhang SG,et al.NDRG2 inhibits migration of breast cancer cells through upregulating PTEN expression[J].Cancer Research on Prevention and Treatment,2017,44(1):5-10.[周晓雷,朱重悦,张世光,等.NDRG2上调PTEN表达抑制乳腺癌细胞迁移的实验[J].肿瘤防治研究,2017,44(1):5-10.]
[16] Gu J,Wang Y,Wang X,et al.Downregulation of lncRNA GAS5 confers tamoxifen resistance by activating miR-222 in breast cancer[J].Cancer Lett,2018,30(434):1-10.