臺灣博碩士論文加值系統

丁基拉草(butachlor)是一種農田除草劑，廣泛的使用在東南亞和台灣的水稻田。研究顯示，丁基拉草可能具有致癌性，但其詳細的致癌機轉仍不清楚。丁基拉草與拉草 (alachlor)在化學結構及性質上非常的相似，拉草是一個已知會導致大鼠鼻咽癌、甲狀腺癌的致癌物質，而且文獻報導拉草會干擾人體的內分泌系統，是一種屬於雌激素類的內分泌干擾物質。因此我們懷疑丁基拉草很可能也是一種致癌物質，或者也是一種內分泌干擾物質。
我們利用細胞生長分析(cell proliferation assay)，發現10、20μM的丁基拉草可以有效的誘使MCF-7和BNL CL2細胞生長分裂。如果把濃度提高，由於丁基拉草cytotoxicity逐漸增加，抵銷掉丁基拉草誘使細胞生長分裂的能力，所以細胞生長的現象反而變得不明顯。為了更進一步瞭解丁基拉草的作用機轉，我們接著評估丁基拉草是否會活化c-jun 和c-fos proto-oncogenes的表現， c-jun和c-fos可以組成AP-1轉錄因子，促進下游基因表現，引起細胞生長及死亡的變化。使用丁基拉草處理30-120分鐘，我們發現無論是MCF-7或BNL CL2的c-jun或c-fos RNA表現都沒有顯著的變化。 把丁基拉草的處理時間延長到36及48小時，MCF-7和BNL CL2的c-jun RNA表現還是沒有顯著的改變。至於c-fos的RNA表現，在MCF-7沒有顯著變化，在BNL CL2則有些微的下降。因此丁基拉草可能不是透過活化AP-1這條MAPK pathway來誘使細胞生長。此外我們也評估丁基拉草是否具有誘導細胞進行轉形(transformation)的能力，所以進行了soft agar assay。從實驗數據中，我們發現單獨使用butachlor或MNNG+butachlor，都無法有效的誘使BNL CL2產生轉形；這個原因可能是因為丁基拉草本身的致癌能力較弱，且經由代謝後，所產生的中間物刺激細胞的時間不夠久，所以還來不及產生轉形所導致。 最後我們使用luciferase reporter assay評估丁基拉草是否是一種內分泌干擾物質。我們的結果顯示40、60μM丁基拉草可以誘導MCF-7細胞luciferase的表現。至於在BNL CL2這株細胞，由於ERα的表現很弱，可能造成ligand和受體親和性不高；或是ligand和受體所形成的complex結合到DNA上的ERE (estrogen response element)序列的能力也是很弱，所以無論是使用E2還是丁基拉草去刺激，luciferase表現都沒有明顯升高。由MCF-7細胞的實驗，丁基拉草很可能也是一種雌激素內分泌干擾物質。
因此總結丁基拉草可能是一種內分泌干擾物質，也可能是一種致癌物質，這必須使用更多的實驗去證明。

Butachlor is a herbicide that has been widely used in rice fields of Southeast Asia and Taiwan. Previous research has shown butachlor could be a carcinogen, but the detailed mechanism is still unclear. Another herbicide, alachlor, is known to cause rat nasopharyngeal carcinoma and thyroid cancer, and has been demonstrated to be an endocrine disrupting chemical (EDC) belonging to an estrogen family. Butachlor and alachlor are very similar in their chemical structure and properties. Therefore, we hypothesized that butachlor might be a carcinogen and EDC.
From the cell proliferation assay, we found that 20μM butachlor could effectively enhance proliferation of both MCF-7 and BNL CL2 cell lines. The cell proliferation became less obvious due to the counter cytotoxic effect of the butachlor if the concentration is higher. To further understand the mechanism of butachlor induced cell proliferation, we examined AP-1 transcription factor regulated pathway. AP-1 is composed of two proto-oncogenes: c-jun and c-fos. Butachlor was used to treat two cell lines, MCF-7 and BNL CL2, for 30 to 120 minutes; there was no significant change in c-jun or c-fos RNA level. If the treatment is lengthened to 36 and 48 hours, still, c-jun RNA level in both MCF-7 and BNL CL2 cell lines showed no difference; however, the level of c-fos RNA was slightly lower in BNL CL2. Therefore, butachlor might not act through AP-1/MAPK pathway to induce cell proliferation. In addition, we would also like to determine if butachlor could induce cell transformation. By performing soft agar assay, we discovered that butachlor alone or with MNNG could not induce BNL CL2 cell line transformation. A possible reason could be the weak carcinogenic ability of butachlor. Lastly, we performed luciferase reporter assay to figure out if butachlor indeed is an EDC. Our results showed that 40 and 60μM of butachlor could induce luciferase expression in MCF-7. As for BNL CL2, whether E2 or butachlor was used, luciferase expression was not significant. A possible explanation could be that the expression of ERα is low, the affinity between the ligand and its receptor was not strong enough, or that between the ligand-receptor complex and estrogen response element (ERE). From MCF-7 experiment, we demonstrated that butachlor could possibly be an estrogen-related EDC. More studies need to be done in the future to confirm the EDC and carcinogenic abilities of butachlor.

中文摘要 ………………………………………………………………………………………… 3
英文摘要 ………………………………………………………………………………………… 4
一、簡介 …………….…………………………………………………………………………… 5
1. Chloroacetamide herbicide ……………………………………………………………… 5
2. Carcinogen ………………………………………………………………………………… 7
3. 雌激素 (estrogen) ………………………………………………………………………… 9
4. 內分泌干擾物質 …………………………………………………………………………… 12
二、目的 ………………………………………………………………………………………… 14
三、材料和方法
Ⅰ. 材料 ………………………………………………………………………………………… 16
Ⅱ. 方法
1. Cell culture ……………………………………………………………………………… 19
2. Cell growth ……………………………………………………………………………… 21
3. RNA extraction ………………………………………………………………………… 22
4. Reverse Transcription ………………………………………………………………… 23
5. 聚合酶連鎖反應 (polymerase chain reaction, PCR) ………………………………… 24
6. 細胞蛋白質萃取及濃度測定 …………………………………………………………… 27
7. 硫酸十二酯鈉聚丙烯醯膠體電泳 (SDSPAGE) ………………………………………… 29
8. 西方點墨法 (western blotting) ………………………………………………………… 31
9. Soft agar assay ………………………………………………………………………… 33
10. Luciferase reporter assay …………………………………………………………… 35
11. Cytotoxicity test ……………………………………………………………………… 37
四、結果 ……………………………………………………………………………………… 39
五、討論 ……………………………………………………………………………………… 44
六、結論 ………………………………………………………………………………………… 50
七、參考文獻 …………………………………………………………………………………… 51
八、實驗圖表 …………………………………………………………………………………… 59
九、附表 ………………………………………………………………………………………… 72

1. Kiely T, Donaldson, D. and Grube, A. Pesticide Industry Sales and Usage: 2000 and 2001 Market Estimates. US Environmental protection Agency2004.
2. Coleman S, Linderman R, Hodgson E, Rose RL. Comparative metabolism of chloroacetamide herbicides and selected metabolites in human and rat liver microsomes. Environ Health Perspect2000 Dec;108(12):1151-7.
3. Matthes B, Boger P. Chloroacetamides affect the plasma membrane. Z Naturforsch C2002 Sep-Oct;57(9-10):843-52.
4. Barbash JE, Thelin GP, Kolpin DW, Gilliom RJ. Major herbicides in ground water: results from the National Water-Quality Assessment. J Environ Qual2001 May-Jun;30(3):831-45.
5. Battaglin WA, Furlong ET, Burkhardt MR, Peter CJ. Occurrence of sulfonylurea, sulfonamide, imidazolinone, and other herbicides in rivers, reservoirs and ground water in the Midwestern United States, 1998. Sci Total Environ2000 Apr 5;248(2-3):123-33.
6. Vargo JD. Determination of sulfonic acid degradates of chloroacetanilide and chloroacetamide herbicides in groundwater by LC/MS/MS. Anal Chem1998 Jul 1;70(13):2699-703.
7. TSENG S-H, CHANG P-C, CHOU S-S. Determination of Butachlor and Pencycuron Residues in Vegetables and Rice: Application of the Macroporous Diatomaceous Earth Column. Journal of Food and Drug Analysis2002;10:127-34.
8. Yu YL, Chen YX, Luo YM, Pan XD, He YF, Wong MH. Rapid degradation of butachlor in wheat rhizosphere soil. Chemosphere2003 Feb;50(6):771-4.
9. EPA U. Chemicals Evaluated for Carcinogenic Potential. US Environmental Protection Agency1999.
10. Wilson AG, Thake DC, Heydens WE, Brewster DW, Hotz KJ. Mode of action of thyroid tumor formation in the male Long-Evans rat administered high doses of alachlor. Fundam Appl Toxicol1996 Sep;33(1):16-23.
11. Genter MB, Burman DM, Dingeldein MW, Clough I, Bolon B. Evolution of alachlor-induced nasal neoplasms in the Long-Evans rat. Toxicol Pathol2000 Nov-Dec;28(6):770-81.
12. Thake DC, Iatropoulos MJ, Hard GC, Hotz KJ, Wang CX, Williams GM, Wilson AG. A study of the mechanism of butachlor-associated gastric neoplasms in Sprague-Dawley rats. Exp Toxicol Pathol1995 May;47(2-3):107-16.
13. Rusiecki JA, Hou L, Lee WJ, Blair A, Dosemeci M, Lubin JH, Bonner M, Samanic C, Hoppin JA, Sandler DP, Alavanja MC. Cancer incidence among pesticide applicators exposed to metolachlor in the Agricultural Health Study. Int J Cancer2006 Jun 15;118(12):3118-23.
14. Wang TC, Lee TC, Lin MF, Lin SY. Induction of sister-chromatid exchanges by pesticides in primary rat tracheal epithelial cells and Chinese hamster ovary cells. Mutat Res1987 Aug;188(4):311-21.
15. Hill AB, Jefferies PR, Quistad GB, Casida JE. Dialkyl- quinoneimine metabolites of chloroacetanilide herbicides induced sister-chromatid exchanges in cultured human lymphocytes. Mutat Res1987;395:159-71.
16. Dearfield KL, McCarroll NE, Protzel A, Stack HF, Jackson MA, Waters MD. A survey of EPA/OPP and open literature on selected pesticide chemicals. II. Mutagenicity and carcinogenicity of selected chloroacetanilides and related compounds. Mutat Res1999 Jul 15;443(1-2):183-221.
17. Hard GC, Iatropoulos MJ, Thake DC, Wheeler D, Tatematsu M, Hagiwara A, Williams GM, Wilson AG. Identity and pathogenesis of stomach tumors in Sprague-Dawley rats associated with the dietary administration of butachlor. Exp Toxicol Pathol1995 May;47(2-3):95-105.
18. Ou YH, Chung PC, Chang YC, Ngo FQ, Hsu KY, Chen FD. Butachlor, a suspected carcinogen, alters growth and transformation characteristics of mouse liver cells. Chem Res Toxicol2000 Dec;13(12):1321-5.
19. Brown MA, Kimmel EC, Casida JE. DNA adduct formation by alachlor metabolites. Life Sci1988;43(25):2087-94.
20. Bonfanti M, Taverna P, Chiappetta L, Villa P, D'Incalci M, Bagnati R, Fanelli R. DNA damage induced by alachlor after in vitro activation by rat hepatocytes. Toxicology1992;72(2):207-19.
21. Taningher M, Terranova MP, Airoldi L, Chiappetta L, Parodi S. Lack of alachlor induced DNA damage as assayed in rodent liver by the alkaline elution test. Toxicology1993 Dec 31;85(2-3):117-22.
22. Jefferies PR, Quistad GB, Casida JE. Dialkylquinonimines validated as in vivo metabolites of alachlor, acetochlor, and metolachlor herbicides in rats. Chem Res Toxicol1998 Apr;11(4):353-9.
23. Singer B, Kusmierek JT. Chemical mutagenesis. Annu Rev Biochem1982;51:655-93.
24. Brown K, Buchmann A, Balmain A. Carcinogen-induced mutations in the mouse c-Ha-ras gene provide evidence of multiple pathways for tumor progression. Proc Natl Acad Sci U S A1990 Jan;87(2):538-42.
25. Iversen OH. Of mice and men: a critical reappraisal of the two-stage theory of carcinogenesis. Crit Rev Oncog1995;6(3-6):357-405.
26. Tennant R. What is a promoter? Environ Health Perspect1999;107:390-91.
27. Barbacid M. ras genes. Annu Rev Biochem1987;56:779-827.
28. Balmain A, Brown K. Oncogene activation in chemical carcinogenesis. Adv Cancer Res1988;51:147-82.
29. Scallen TJ, Noland BJ, Gavey KL, Bass NM, Ockner RK, Chanderbhan R, Vahouny GV. Sterol carrier protein 2 and fatty acid-binding protein. Separate and distinct physiological functions. J Biol Chem1985 Apr 25;260(8):4733-9.
30. Osawa Y, Higashiyama T, Shimizu Y, Yarborough C. Multiple functions of aromatase and the active site structure; aromatase is the placental estrogen 2-hydroxylase. J Steroid Biochem Mol Biol1993 Mar;44(4-6):469-80.
31. Gruber DM, Huber JC. Tissue specificity: the clinical importance of steroid metabolites in hormone replacement therapy. Maturitas2001 Jan 31;37(3):151-7.
32. Liehr JG. Is estradiol a genotoxic mutagenic carcinogen? Endocr Rev2000 Feb;21(1):40-54.
33. Paganini-Hill A, Dworsky R, Krauss RM. Hormone replacement therapy, hormone levels, and lipoprotein cholesterol concentrations in elderly women. Am J Obstet Gynecol1996 Mar;174(3):897-902.
34. Gruber CJ, Tschugguel W, Schneeberger C, Huber JC. Production and actions of estrogens. N Engl J Med2002 Jan 31;346(5):340-52.
35. Green S, Walter P, Kumar V, Krust A, Bornert JM, Argos P, Chambon P. Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A. Nature1986 Mar 13-19;320(6058):134-9.
36. Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA. Cloning of a novel receptor expressed in rat prostate and ovary. Proc Natl Acad Sci U S A1996 Jun 11;93(12):5925-30.
37. Witkowska HE, Carlquist M, Engstrom O, Carlsson B, Bonn T, Gustafsson JA, Shackleton CH. Characterization of bacterially expressed rat estrogen receptor beta ligand binding domain by mass spectrometry: structural comparison with estrogen receptor alpha. Steroids1997 Aug-Sep;62(8-9):621-31.
38. Kuiper GG, Carlsson B, Grandien K, Enmark E, Haggblad J, Nilsson S, Gustafsson JA. Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. Endocrinology1997 Mar;138(3):863-70.
39. Enmark E, Pelto-Huikko M, Grandien K, Lagercrantz S, Lagercrantz J, Fried G, Nordenskjold M, Gustafsson JA. Human estrogen receptor beta-gene structure, chromosomal localization, and expression pattern. J Clin Endocrinol Metab1997 Dec;82(12):4258-65.
40. Pettersson K, Grandien K, Kuiper GG, Gustafsson JA. Mouse estrogen receptor beta forms estrogen response element-binding heterodimers with estrogen receptor alpha. Mol Endocrinol1997 Sep;11(10):1486-96.
41. Webb P, Lopez GN, Uht RM, Kushner PJ. Tamoxifen activation of the estrogen receptor/AP-1 pathway: potential origin for the cell-specific estrogen-like effects of antiestrogens. Mol Endocrinol1995 Apr;9(4):443-56.
42. Porter JC. Proceedings: Hormonal regulation of breast development and activity. J Invest Dermatol1974 Jul;63(1):85-92.
43. Yue W, Santen RJ, Wang JP, Hamilton CJ, Demers LM. Aromatase within the breast. Endocr Relat Cancer1999 Jun;6(2):157-64.
44. Fox JE, Starcevic M, Jones PE, Burow ME, McLachlan JA. Phytoestrogen signaling and symbiotic gene activation are disrupted by endocrine-disrupting chemicals. Environ Health Perspect2004 May;112(6):672-7.
45. Longnecker MP, Rogan WJ, Lucier G. The human health effects of DDT (dichlorodiphenyltrichloroethane) and PCBS (polychlorinated biphenyls) and an overview of organochlorines in public health. Annu Rev Public Health1997;18:211-44.
46. Kelce WR, Stone CR, Laws SC, Gray LE, Kemppainen JA, Wilson EM. Persistent DDT metabolite p,p'-DDE is a potent androgen receptor antagonist. Nature1995 Jun 15;375(6532):581-5.
47. Zacharewski T. Identification and assessment of endocrine disruptors: limitations of in vivo and in vitro assays. Environ Health Perspect1998 Apr;106 Suppl 2:577-82.
48. Cheek AO, Vonier PM, Oberdorster E, Burow BC, McLachlan JA. Environmental signaling: a biological context for endocrine disruption. Environ Health Perspect1998 Feb;106 Suppl 1:5-10.
49. Andersen ME, Conolly RB, Faustman EM, Kavlock RJ, Portier CJ, Sheehan DM, Wier PJ, Ziese L. Quantitative mechanistically based dose-response modeling with endocrine-active compounds. Environ Health Perspect1999 Aug;107 Suppl 4:631-8.
50. Roy D, Palangat M, Chen CW, Thomas RD, Colerangle J, Atkinson A, Yan ZJ. Biochemical and molecular changes at the cellular level in response to exposure to environmental estrogen-like chemicals. J Toxicol Environ Health1997 Jan;50(1):1-29.
51. McLachlan JA. Environmental signaling: what embryos and evolution teach us about endocrine disrupting chemicals. Endocr Rev2001 Jun;22(3):319-41.
52. Tyler CR, Jobling S, Sumpter JP. Endocrine disruption in wildlife: a critical review of the evidence. Crit Rev Toxicol1998 Jul;28(4):319-61.
53. Danzo BJ. Environmental xenobiotics may disrupt normal endocrine function by interfering with the binding of physiological ligands to steroid receptors and binding proteins. Environ Health Perspect1997 Mar;105(3):294-301.
54. Facemire CF, Gross TS, Guillette LJ, Jr. Reproductive impairment in the Florida panther: nature or nurture? Environ Health Perspect1995 May;103 Suppl 4:79-86.
55. Welch RM, Levin W, Conney AH. Estrogenic action of DDT and its analogs. Toxicol Appl Pharmacol1969 Mar;14(2):358-67.
56. Cummings AM. Methoxychlor as a model for environmental estrogens. Crit Rev Toxicol1997 Jul;27(4):367-79.
57. Coosen R, van Velsen FL. Effects of the beta-isomer of hexachlorocyclohexane on estrogen-sensitive human mammary tumor cells. Toxicol Appl Pharmacol1989 Nov;101(2):310-8.
58. Soto AM, Sonnenschein C, Chung KL, Fernandez MF, Olea N, Serrano FO. The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants. Environ Health Perspect1995 Oct;103 Suppl 7:113-22.
59. Garey J, Wolff MS. Estrogenic and antiprogestagenic activities of pyrethroid insecticides. Biochem Biophys Res Commun1998 Oct 29;251(3):855-9.
60. Burow ME, Tang Y, Collins-Burow BM, Krajewski S, Reed JC, McLachlan JA, Beckman BS. Effects of environmental estrogens on tumor necrosis factor alpha-mediated apoptosis in MCF-7 cells. Carcinogenesis1999 Nov;20(11):2057-61.
61. Lee C, Ryu J, Park S, Choi K, Jeon S, Na J, Rhee D. Effects of alachlor on the early development and induction of estrogen-responsive genes in medaka, Oryzias Iatipes. Organohalogen Compounds2004;66:2993-99.
62. Mueller SO. Xenoestrogens: mechanisms of action and detection methods. Anal Bioanal Chem2004 Feb;378(3):582-7.
63. Bolger R, Wiese TE, Ervin K, Nestich S, Checovich W. Rapid screening of environmental chemicals for estrogen receptor binding capacity. Environ Health Perspect1998 Sep;106(9):551-7.
64. Nikov GN, Hopkins NE, Boue S, Alworth WL. Interactions of dietary estrogens with human estrogen receptors and the effect on estrogen receptor-estrogen response element complex formation. Environ Health Perspect2000 Sep;108(9):867-72.
65. Boyer M, Poujol N, Margeat E, Royer CA. Quantitative characterization of the interaction between purified human estrogen receptor alpha and DNA using fluorescence anisotropy. Nucleic Acids Res2000 Jul 1;28(13):2494-502.
66. Mueller SO, Kling M, Arifin Firzani P, Mecky A, Duranti E, Shields-Botella J, Delansorne R, Broschard T, Kramer PJ. Activation of estrogen receptor alpha and ERbeta by 4-methylbenzylidene-camphor in human and rat cells: comparison with phyto- and xenoestrogens. Toxicol Lett2003 Apr 30;142(1-2):89-101.
67. Shelby MD, Newbold RR, Tully DB, Chae K, Davis VL. Assessing environmental chemicals for estrogenicity using a combination of in vitro and in vivo assays. Environ Health Perspect1996 Dec;104(12):1296-300.
68. Sanderson JT, Boerma J, Lansbergen GW, van den Berg M. Induction and inhibition of aromatase (CYP19) activity by various classes of pesticides in H295R human adrenocortical carcinoma cells. Toxicol Appl Pharmacol2002 Jul 1;182(1):44-54.
69. Ando S, Sirianni R, Forastieri P, Casaburi I, Lanzino M, Rago V, Giordano F, Giordano C, Carpino A, Pezzi V. Aromatase expression in prepuberal Sertoli cells: effect of thyroid hormone. Mol Cell Endocrinol2001 Jun 10;178(1-2):11-21.
70. Gray LE, Jr., Ostby J, Wilson V, Lambright C, Bobseine K, Hartig P, Hotchkiss A, Wolf C, Furr J, Price M, Parks L, Cooper RL, Stoker TE, Laws SC, Degitz SJ, Jensen KM, Kahl MD, Korte JJ, Makynen EA, Tietge JE, Ankley GT. Xenoendocrine disrupters-tiered screening and testing: filling key data gaps. Toxicology2002 Dec 27;181-182:371-82.
71. Rotchell JM, Ostrander GK. Molecular markers of endocrine disruption in aquatic organisms. J Toxicol Environ Health B Crit Rev2003 Sep-Oct;6(5):453-96.
72. Purdom CE, Hardiman PA, Bye VVJ, Eno NC, Tyler CR, Sumpter JP. Estrogenic Effects of Effluents from Sewage Treatment Works. Chemistry and Ecology1994;8(4):275 - 85.
73. Routledge EJ, Sheahan D, Desbrow C, Brighty GC, Waldock M, Sumpter JP. Identification of Estrogenic Chemicals in STW Effluent. 2. In Vivo Responses in Trout and Roach. Environmental Science & Technology1998;32(11):1559-65.
74. Driever W, Solnica-Krezel L, Schier AF, Neuhauss SC, Malicki J, Stemple DL, Stainier DY, Zwartkruis F, Abdelilah S, Rangini Z, Belak J, Boggs C. A genetic screen for mutations affecting embryogenesis in zebrafish. Development1996 Dec;123:37-46.
75. Talbot WS, Trevarrow B, Halpern ME, Melby AE, Farr G, Postlethwait JH, Jowett T, Kimmel CB, Kimelman D. A homeobox gene essential for zebrafish notochord development. Nature1995 Nov 9;378(6553):150-7.
76. Stainier DY, Weinstein BM, Detrich HW, 3rd, Zon LI, Fishman MC. Cloche, an early acting zebrafish gene, is required by both the endothelial and hematopoietic lineages. Development1995 Oct;121(10):3141-50.
77. Abdelilah S, Solnica-Krezel L, Stainier DY, Driever W. Implications for dorsoventral axis determination from the zebrafish mutation janus. Nature1994 Aug 11;370(6489):468-71.
78. Oppen-Berntsen DO, Gram-Jensen E, Walther BT. Zona radiata proteins are synthesized by rainbow trout (Oncorhynchus mykiss) hepatocytes in response to oestradiol-17 beta. J Endocrinol1992 Nov;135(2):293-302.
79. Tsai CL, Wang LH, Fang LS. Estradiol and para-chlorophenylalanine downregulate the expression of brain aromatase and estrogen receptor-alpha mRNA during the critical period of feminization in tilapia (Oreochromis mossambicus). Neuroendocrinology2001 Nov;74(5):325-34.
80. Rocha Monteiro PR, Reis-Henriques MA, Coimbra J. Polycyclic aromatic hydrocarbons inhibit in vitro ovarian steroidogenesis in the flounder (Platichthys flesus L.). Aquat Toxicol2000 Apr 1;48(4):549-59.
81. Kishida M, Callard GV. Distinct cytochrome P450 aromatase isoforms in zebrafish (Danio rerio) brain and ovary are differentially programmed and estrogen regulated during early development. Endocrinology2001 Feb;142(2):740-50.
82. Kishida M, McLellan M, Miranda JA, Callard GV. Estrogen and xenoestrogens upregulate the brain aromatase isoform (P450aromB) and perturb markers of early development in zebrafish (Danio rerio). Comp Biochem Physiol B Biochem Mol Biol2001 Jun;129(2-3):261-8.
83. Sampath Kumar R, Ijiri S, Trant JM. Changes in the expression of genes encoding steroidogenic enzymes in the channel catfish (Ictalurus punctatus) ovary throughout a reproductive cycle. Biol Reprod2000 Dec;63(6):1676-82.
84. Bernstein LR, Colburn NH. AP1/jun function is differentially induced in promotion-sensitive and resistant JB6 cells. Science1989 May 5;244(4904):566-9.
85. Carter T, James C, Chan E, Greene B. Induction of integrated adenovirus E1A and E1B genes in transformed human cells by phorbol ester tumor promoters. Cancer Res1987 Feb 1;47(3):803-8.
86. Mangelsdorf DJ, Thummel C, Beato M, Herrlich P, Schutz G, Umesono K, Blumberg B, Kastner P, Mark M, Chambon P, Evans RM. The nuclear receptor superfamily: the second decade. Cell1995 Dec 15;83(6):835-9.
87. Beato M, Herrlich P, Schutz G. Steroid hormone receptors: many actors in search of a plot. Cell1995 Dec 15;83(6):851-7.
88. Le Goff P, Montano MM, Schodin DJ, Katzenellenbogen BS. Phosphorylation of the human estrogen receptor. Identification of hormone-regulated sites and examination of their influence on transcriptional activity. J Biol Chem1994 Feb 11;269(6):4458-66.
89. Levin ER. Cellular Functions of the Plasma Membrane Estrogen Receptor. Trends Endocrinol Metab1999 Nov;10(9):374-7.
90. Kelly MJ, Wagner EJ. Estrogen Modulation of G-protein-coupled Receptors. Trends Endocrinol Metab1999 Nov;10(9):369-74.
91. Castoria G, Barone MV, Di Domenico M, Bilancio A, Ametrano D, Migliaccio A, Auricchio F. Non-transcriptional action of oestradiol and progestin triggers DNA synthesis. EMBO J1999 May 4;18(9):2500-10.
92. Razandi M, Pedram A, Levin ER. Plasma membrane estrogen receptors signal to antiapoptosis in breast cancer. Mol Endocrinol2000 Sep;14(9):1434-47.
93. Zacharewski T. Identification and assessment of endocrine disruptors: limitations of in vivo and in vitro assays. Environ Health Perspect1998;106(Suppl 2):577–82.
94. Heydens WF. Summary of toxicology studies with alachlor. J Pesticide Sci1999;24:75-82.
95. Klotz DM, Beckman BS, Hill SM, McLachlan JA, Walters MR, Arnold SF. Identification of environmental chemicals with estrogenic activity using a combination of in vitro assays. Environ Health Perspect1996 Oct;104(10):1084-9.
96. Soto AM, Chung KL, Sonnenschein C. The pesticides endosulfan, toxaphene, and dieldrin have estrogenic effects on human estrogen-sensitive cells. Environ Health Perspect1994 Apr;102(4):380-3.
97. Arts J, Herr I, Lansink M, Angel P, Kooistra T. Cell-type specific DNA-protein interactions at the tissue-type plasminogen activator promoter in human endothelial and HeLa cells in vivo and in vitro. Nucleic Acids Res1997 Jan 15;25(2):311-7.
98. Diane M. Klotz BSB, McLachlan, Marian Walters, Steven F. Arnold. Identification of Environmental Chemicals with Estrogenic Activity Using a Combination of In Vitro Assays. Environmental Health Perspectives1996 Number 10, October 1996;104:1084-9.
99. Albanito L, Lappano R, Madeo A, Chimento A, Prossnitz ER, Cappello AR, Dolce V, Abonante S, Pezzi V, Maggiolini M. G-Protein–Coupled Receptor 30 and Estrogen Receptor-α Are Involved in the Proliferative Effects Induced by Atrazine in Ovarian Cancer Cells. Environmental Health Perspectives2008;116(1Department of Pharmaco-Biology, University of Calabria, Rende, Italy; 2Department of Cell Biology and Physiology and Cancer
Research and Treatment Center, University of New Mexico, Albuquerque, New Mexico, USA):1648-55.
100. Aube M, Larochelle C, Ayotte P. 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (p,p'-DDE) disrupts the estrogen-androgen balance regulating the growth of hormone-dependent breast cancer cells. Breast Cancer Research2008;10:12.
101. WILSON AGE, TAKEI AS. Summary of Toxicology Studies with Butachlor. 1999(Monsanto Company).