牛秀瓏 1 , 王奕飛 2 , 孫奕 1 , 王越 1,3
  • 1.武警醫(yī)學(xué)院免疫學(xué)教研室（天津 300162）;;
  • 2.武警醫(yī)學(xué)院科研部學(xué)報(bào)編輯部（天津 300162）;;
  • 3.武警醫(yī)學(xué)院天津市職業(yè)與環(huán)境危害生物標(biāo)志物重點(diǎn)實(shí)驗(yàn)室（天津 300162）;

目的構(gòu)建雌激素受體（ER）亞型ERα/ERβ不同表達(dá)狀態(tài)的人乳腺癌細(xì)胞株MCF-7，觀察其生物學(xué)特性。方法采用RNA干擾技術(shù)沉默ERα或ERβ基因表達(dá)，獲得不同ERα或ERβ表達(dá)狀態(tài)的MCF-7細(xì)胞株。 運(yùn)用MTT法、流式細(xì)胞術(shù)、RT-PCR法、雙層軟瓊脂集落形成實(shí)驗(yàn)和體外細(xì)胞黏附實(shí)驗(yàn)方法檢測(cè)細(xì)胞增殖、凋亡、體外成瘤和黏附能力。結(jié)果構(gòu)建穩(wěn)定表達(dá)ERαlow/ERβhigh或ERαhigh/ERβlow的MCF-7細(xì)胞株。 ERα基因沉默后，細(xì)胞生長(zhǎng)減慢，受阻于G0～G1期，細(xì)胞凋亡增加，體外成瘤能力和黏附能力減弱； ERβ基因沉默后，細(xì)胞生長(zhǎng)加快，S期細(xì)胞比例增加，細(xì)胞凋亡受到抑制，體外成瘤能力增強(qiáng)，而黏附特性無(wú)變化。 結(jié)論ERα基因沉默可抑制腫瘤的形成； 而ERβ基因沉默可促進(jìn)腫瘤生長(zhǎng)、侵襲和轉(zhuǎn)移。 調(diào)節(jié)ERα/ERβ表達(dá)狀態(tài)可能會(huì)成為一種有效的乳腺癌治療手段。

引用本文： 牛秀瓏,王奕飛,孫奕,王越. 雌激素受體亞型對(duì)人乳腺癌細(xì)胞株MCF-7生物學(xué)特性的影響. 中國(guó)普外基礎(chǔ)與臨床雜志, 2011, 18(7): 711-716. doi: 復(fù)制

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2. Welboren WJ, Sweep FC, Span PN, et al. Genomic actions of estrogen receptor alpha: what are the targets and how are they regulated? [J]. Endocr Relat Cancer, 2009, 16(4): 10731089.
3. Paech K, Webb P, Kuiper GG, et al. Differential ligand activation of estrogen receptors ERalpha and ERbeta at AP1 sites [J]. Science, 1997, 277(5331): 15081510.
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11. Strom A, Hartman J, Foster JS, et al. Estrogen receptor beta inhibits 17 betaestradiolstimulated proliferation of the breast cancer cell line T47D [J]. Proc Natl Acad Sci USA, 2004, 101(6): 15661571.
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15. Tonetti DA, Rubenstein R, DeLeon M, et al. Stable transfection of an estrogen receptor beta cDNA isoform into MDAMB231 breast cancer cells [J]. J Steroid Biochem Mol Biol, 2003, 87(1): 4755.
16. Hou YF, Yuan ST, Li HC, et al. ERbeta exerts multiple stimulative effects on human breast carcinoma cells [J]. Oncogene, 2004, 23(34): 57995806.
17. 侯意楓， 袁勝濤， 李鶴成， 等. 雌激素受體β對(duì)人乳腺癌細(xì)胞株生物學(xué)特性的影響 [J]. 中華腫瘤雜志， 2005, 27(7): 389392.
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19. 吳敏， 吳靜， 徐玥， 等. 小RNA干擾PDK1抑制乳腺腫瘤細(xì)胞侵襲 [J]. 中國(guó)腫瘤臨床, 2009, 36(12): 697700.
20. 董林， 何曉東， 陶紹能， 等. 甲氨蝶呤對(duì)映體誘導(dǎo)的肺癌A549耐藥細(xì)胞株中VEGF及其受體表達(dá)差異的研究 [J]. 腫瘤, 2009, 29(5): 404408.
21. Myler HA, West JL. Heparanase and platelet factor4 induce smooth muscle cell proliferation and migration via bFGF release from the ECM [J]. J Biochem, 2002, 131(6): 913922.
22. 高德宗， 高華， 郭秀會(huì)， 等. 他莫昔芬與γ干擾素聯(lián)合抗人乳腺癌細(xì)胞株的作用及其機(jī)制研究 [J]. 中國(guó)普通外科雜志， 2007， 16(11)： 10761080.
23. 高德宗， 孫靖中， 高華， 等. γ干擾素增強(qiáng)他莫昔芬抗乳腺癌作用的體外研究 [J]. 中國(guó)腫瘤生物治療雜志， 2006， 13(1)： 4549.
24. Pedicini M, Barrens F, Clancy T, et al. Combining network modeling and gene expression microarray analysis to explore the dynamics of Th1 and Th2 cell regulation [J]. PLoS Comput Biol, 2010, 6(12): e1001032.
25. 孫桂明， 李新煥， 王寶中， 等. 維甲酸聯(lián)合他莫昔芬對(duì)人乳腺癌細(xì)胞株作用的研究 [J]. 中華腫瘤防治雜志， 2010， 17(18)： 14251428.
  1. 1. Nangaku M. Chronic hypoxia and tubulointerstitial injury: a final common pathway to endstage renal failure [J]. J Am Soc Nephrol, 2006, 17(1): 1725.
  2. 2. Welboren WJ, Sweep FC, Span PN, et al. Genomic actions of estrogen receptor alpha: what are the targets and how are they regulated? [J]. Endocr Relat Cancer, 2009, 16(4): 10731089.
  3. 3. Paech K, Webb P, Kuiper GG, et al. Differential ligand activation of estrogen receptors ERalpha and ERbeta at AP1 sites [J]. Science, 1997, 277(5331): 15081510.
  4. 4. Hayashi SI, Eguchi H, Tanimoto K, et al. The expression and function of estrogen receptor alpha and beta in human breast cancer and its clinical application [J]. Endocr Relat Cancer, 2003, 10(2): 193202.
  5. 5. Meacham WD, Antoon JW, Burow ME, et al. Sphingolipids as determinants of apoptosis and chemoresistance in the MCF7 cell model system [J]. Exp Biol Med (Maywood), 2009， 234(11): 1253 1263.
  6. 6. Speirs V. Oestrogen receptor in breast cancer: good, bad or still too early to tell [J]. J Pathol, 2002, 197(2)： 143147.
  7. 7. Zhao C, DahlmanWright K, Gustafsson JA. Estrogen signaling via estrogen receptor beta [J]. J Biol Chem, 2010, 285(51): 3957539579.
  8. 8. Drummond AE, Fuller PJ. The importance of ERbeta signalling in the ovary [J]. J Endocrinol, 2010, 205(1): 1523.
  9. 9. Roger P, Sahla ME, Makela S, et al. Decreased expression of estrogen receptor beta protein in proliferative preinvasive mammary tumors [J]. Cancer Res, 2001, 61(6): 25372541.
  10. 10. Paruthiyil S, Parmar H, Kerekatte V, et al. Estrogen receptor beta inhibits human breast cancer cell proliferation and tumor formation by causing a G2 cell cycle arrest [J]. Cancer Res, 2004, 64(1): 423428.
  11. 11. Strom A, Hartman J, Foster JS, et al. Estrogen receptor beta inhibits 17 betaestradiolstimulated proliferation of the breast cancer cell line T47D [J]. Proc Natl Acad Sci USA, 2004, 101(6): 15661571.
  12. 12. Helguero LA, Faulds MH, Gustafsson JA, et al. Estrogen receptors alfa (ERalpha) and beta (ERbeta) differentially regulate proliferation and apoptosis of the normal murine mammary epithelial cell line HC11 [J]. Oncogene, 2005, 24(44): 66056616.
  13. 13. Chen L, Qiu J, Yang C, et al. Identification of a novel estrogen receptor beta1 binding partner, inhibitor of differentiation1, and role of ERbeta1 in human breast cancer cells [J]. Cancer Lett, 2009, 278(2): 210219．.
  14. 14. Grober OM, Mutarelli M, Giurato G, et al. Global analysis of estrogen receptor beta binding to breast cancer cell genome reveals an extensive interplay with estrogen receptor alpha for target gene regulation [J]. BMC Genomics, 2011, 14(12): 3652.
  15. 15. Tonetti DA, Rubenstein R, DeLeon M, et al. Stable transfection of an estrogen receptor beta cDNA isoform into MDAMB231 breast cancer cells [J]. J Steroid Biochem Mol Biol, 2003, 87(1): 4755.
  16. 16. Hou YF, Yuan ST, Li HC, et al. ERbeta exerts multiple stimulative effects on human breast carcinoma cells [J]. Oncogene, 2004, 23(34): 57995806.
  17. 17. 侯意楓， 袁勝濤， 李鶴成， 等. 雌激素受體β對(duì)人乳腺癌細(xì)胞株生物學(xué)特性的影響 [J]. 中華腫瘤雜志， 2005, 27(7): 389392.
  18. 18. 于冰， 孫治君. 凋亡相關(guān)基因bcl2、bax、bad與乳腺癌 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2007, 14(6): 739742.
  19. 19. 吳敏， 吳靜， 徐玥， 等. 小RNA干擾PDK1抑制乳腺腫瘤細(xì)胞侵襲 [J]. 中國(guó)腫瘤臨床, 2009, 36(12): 697700.
  20. 20. 董林， 何曉東， 陶紹能， 等. 甲氨蝶呤對(duì)映體誘導(dǎo)的肺癌A549耐藥細(xì)胞株中VEGF及其受體表達(dá)差異的研究 [J]. 腫瘤, 2009, 29(5): 404408.
  21. 21. Myler HA, West JL. Heparanase and platelet factor4 induce smooth muscle cell proliferation and migration via bFGF release from the ECM [J]. J Biochem, 2002, 131(6): 913922.
  22. 22. 高德宗， 高華， 郭秀會(huì)， 等. 他莫昔芬與γ干擾素聯(lián)合抗人乳腺癌細(xì)胞株的作用及其機(jī)制研究 [J]. 中國(guó)普通外科雜志， 2007， 16(11)： 10761080.
  23. 23. 高德宗， 孫靖中， 高華， 等. γ干擾素增強(qiáng)他莫昔芬抗乳腺癌作用的體外研究 [J]. 中國(guó)腫瘤生物治療雜志， 2006， 13(1)： 4549.
  24. 24. Pedicini M, Barrens F, Clancy T, et al. Combining network modeling and gene expression microarray analysis to explore the dynamics of Th1 and Th2 cell regulation [J]. PLoS Comput Biol, 2010, 6(12): e1001032.
  25. 25. 孫桂明， 李新煥， 王寶中， 等. 維甲酸聯(lián)合他莫昔芬對(duì)人乳腺癌細(xì)胞株作用的研究 [J]. 中華腫瘤防治雜志， 2010， 17(18)： 14251428.