Hsp90抑制剂WK-88-1对人乳腺癌细胞迁移与侵袭的影响
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摘要
目的探讨热休克蛋白90(Hsp90)抑制剂WK-88-1对人乳腺癌细胞迁移与侵袭能力的影响。方法 MTT实验检测不同浓度的WK-88-1(1. 562 5,3. 125,6. 25,12. 5,25,50μmol/L)对人乳腺癌MDA-MB-231和MCF-7细胞存活率的影响;集落克隆实验检测不同浓度的WK-88-1(0. 1,0. 3,0. 5μmol/L)对人乳腺癌MDA-MB-231和MCF-7细胞集落形成的影响;细胞划痕实验和Transwell小室法检测WK-88-1对人乳腺癌MDA-MB-231和MCF-7细胞迁移和侵袭的影响;免疫印迹法检测WK-88-1对Hsp90顾客蛋白缺氧诱导因子-1α(HIF-1α)和基质金属蛋白酶-9(MMP-9)蛋白表达的影响。结果 WK-88-1具有抑制MDA-MB-231和MCF-7细胞增殖的作用,且呈现浓度和时间依赖性(P<0.05)。MDA-MB-231和MCF-7经不同浓度的WK-88-1处理后,癌细胞的迁移和侵袭能力显著下降(P<0.05)。免疫印迹法结果显示,随着WK-88-1给药浓度(12. 5,25,50μmol/L)的增加,细胞内的HIF-1α和MMP-9蛋白的表达均下调。结论非苯醌格尔德霉素WK-88-1能够显著抑制人乳腺癌细胞迁移与侵袭,其机制可能与下调Hsp90的顾客蛋白HIF-1α和MMP-9有关。
Objective To investigate the effect of WK-88-1,a heat shock protein 90(Hsp90) inhibitor,on the migration and invasion of human breast cancer cells. Methods MTT assay was used to detect the survival rate of human breast cancer MDA-MB-231 and MCF-7 cells after treated with WK-88-1(1. 562 5,3. 125,6. 25,12. 5,25,50 μmol/L). The effects of WK-88-1(0. 1,0. 3,0. 5 μmol/L) on the colony formation of MDA-MB-231 and MCF-7 cells were observed by colony formation experiment. The effect of WK-88-1 on migration and invasion abilities of breast cancer cells were analyzed using wound healing assay and transwell assay. The effects of WK-88-1 on the expression levels of Hsp90 clinent proteins HIF-1α and MMP-9 were detected using Western blotting. Results WK-88-1 exhibited anti-proliferation activities against MDA-MB-231 and MCF-7 cells in a dose-and time-dependent manner(P<0.05). After treatment with different concentrations of WK-88-1,the breast cancer cell migration and invasion abilities significantly decreased(P <0. 05). Western blotting results showed that expression levels of HIF-1α and MMP-9 down-regulated by WK-88-1 in a concentrationdependent manner(12. 5,25,50 μmol/L). Conclusion Non-benzoquinone geldanamycin WK-88-1 significantly inhibit the migration and invasion of human breast cancer cells,which may be related to the down-regulated expression of Hsp90 client proteins HIF-1α and MMP-9.
Objective To investigate the effect of WK-88-1,a heat shock protein 90(Hsp90) inhibitor,on the migration and invasion of human breast cancer cells. Methods MTT assay was used to detect the survival rate of human breast cancer MDA-MB-231 and MCF-7 cells after treated with WK-88-1(1. 562 5,3. 125,6. 25,12. 5,25,50 μmol/L). The effects of WK-88-1(0. 1,0. 3,0. 5 μmol/L) on the colony formation of MDA-MB-231 and MCF-7 cells were observed by colony formation experiment. The effect of WK-88-1 on migration and invasion abilities of breast cancer cells were analyzed using wound healing assay and transwell assay. The effects of WK-88-1 on the expression levels of Hsp90 clinent proteins HIF-1α and MMP-9 were detected using Western blotting. Results WK-88-1 exhibited anti-proliferation activities against MDA-MB-231 and MCF-7 cells in a dose-and time-dependent manner(P<0.05). After treatment with different concentrations of WK-88-1,the breast cancer cell migration and invasion abilities significantly decreased(P <0. 05). Western blotting results showed that expression levels of HIF-1α and MMP-9 down-regulated by WK-88-1 in a concentrationdependent manner(12. 5,25,50 μmol/L). Conclusion Non-benzoquinone geldanamycin WK-88-1 significantly inhibit the migration and invasion of human breast cancer cells,which may be related to the down-regulated expression of Hsp90 client proteins HIF-1α and MMP-9.
引文
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[14] Hyslop T,Michael Y,Avery T,et al. Population and target considerations for triple-negative breast cancer clinical trials[J]. Biomark Med,2013,7(1):11-21.
[15]胡瑛,李宝兰.缺氧诱导因子-1α、2α在恶性肿瘤中不同作用的研究进展[J].癌症进展,2014,12(2):122-125.
[16] Nagaraju GP,Bramhachari PV,Raghu G,et al. Hypoxia inducible factor-1α:Its role in colorectal carcinogenesis and metastasis[J]. Cancer Lett,2015,366(1):11-18.
[17] Chu CY,Jin YT,Zhang W,et al. CAIX is upregulated in CoCl2-induced hypoxia and associated with cell invasive potential and a poor prognosis of breast cancer[J]. Intern J Oncol,2016,48(1):271-280.
[18] Fan SH,Wang YY,Lu J,et al. CERS2 suppresses tumor cell invasion and is associated with decreased V-ATPase and MMP-2/MMP-9 activities in breast cancer[J]. J Cellular Biochem,2015,116(4):502-513.
[19]姚广裕,陈路嘉,胡晓磊,等. MT1-MMP在乳腺癌中的独特表达模式[J].南方医科大学学报,2016,36(1):94-97.
[2] Streb J,Glanowska I,Streb A,et al. The relationship between breast cancer treatment,tumour type and vitamin D level in preand postmenopausal women[J]. Neuro Endocrinol Lett,2017,38(6):437-440.
[3]郑莹,吴春晓,张敏璐.乳腺癌在中国的流行状况和疾病特征[J].中国癌症杂志,2013,23(8):561-569.
[4] Lee HJ,Seo NJ,Jeong SJ,et al. Oral administration of penta-Ogalloyl-beta-D-glucose suppresses triple-negative breast cancer xenograft growth and metastasis in strong association with JAKISTAT3 inhibition[J]. Carcinogenesis,2011,32(6):804-811.
[5] Whitesell L,Lindquist SL. Hsp90 and the chaperoning of cancer[J]. Nat Rev Cancer,2005,5(10):761-772.
[6] Pearl LH,Prodromou C,Workman P. The Hsp90 molecular charperone:an open and shut case for treatment[J]. Biochem J,2008,410(3):439-453.
[7] Sankhala KK,Mita MM,Mita AC,et al. Heat shock proteins:a potential anticancer target[J]. Curr Drug Targets,2011,12(14):2001-2008.
[8] Kim W,Lee J,Lee D,et al. Mutasynthesis of geldanamycin by the disruption of a gene producing starter unit:generation of structural diversity at the benzoquinone ring[J]. Chem Bio Chem,2007,8(13):1491-1494.
[9] Jang WJ,Jung SK,Kang JS,et al. Anti-tumor acitivity of WK-88-1,a novel geldanamycin derivative,in gefitinib-resistant nonsmall cell lung cancers with Met amplification[J]. Cancer Sci,2014,105(10):1245-1253.
[10] Hong YS,Jang WJ,Chun KS,et al. Hsp90 inhibition by WK-88-1 potently suppresses the growth of geftinib-resistant H1975cells harboring the T790M mutation in EGFR[J]. Oncol Rep,2014,31:2619-2624.
[11] Zhao YR,Li HM,Zhu M,et al. Non-benzoquinone geldanamycin analog,WK-88-1,induces apoptosis in human breast cancer cell lines[J]. J Microbiol Biotechnol,2018,28(4):542-550.
[12]苗健,马涛,李楠,等.厚朴酚葡萄糖苷对人乳腺癌细胞增殖、侵袭、迁移的影响[J].蚌埠医学院学报,2018,43(5):567-572.
[13] Shastry M,Yardley D. Updates in the treatment of basal/triplenegative breast cancer[J]. Curr Opin Obstet Gynecol,2013,25(1):40-48.
[14] Hyslop T,Michael Y,Avery T,et al. Population and target considerations for triple-negative breast cancer clinical trials[J]. Biomark Med,2013,7(1):11-21.
[15]胡瑛,李宝兰.缺氧诱导因子-1α、2α在恶性肿瘤中不同作用的研究进展[J].癌症进展,2014,12(2):122-125.
[16] Nagaraju GP,Bramhachari PV,Raghu G,et al. Hypoxia inducible factor-1α:Its role in colorectal carcinogenesis and metastasis[J]. Cancer Lett,2015,366(1):11-18.
[17] Chu CY,Jin YT,Zhang W,et al. CAIX is upregulated in CoCl2-induced hypoxia and associated with cell invasive potential and a poor prognosis of breast cancer[J]. Intern J Oncol,2016,48(1):271-280.
[18] Fan SH,Wang YY,Lu J,et al. CERS2 suppresses tumor cell invasion and is associated with decreased V-ATPase and MMP-2/MMP-9 activities in breast cancer[J]. J Cellular Biochem,2015,116(4):502-513.
[19]姚广裕,陈路嘉,胡晓磊,等. MT1-MMP在乳腺癌中的独特表达模式[J].南方医科大学学报,2016,36(1):94-97.