Alterations in growth factor signaling pathways frequently accompany the development and maintenance of epithelial neoplasia. Transforming growth factor a (TGFα) and its epidermal growth factor receptor have been thought to play an especially important role in epithelial neoplasia. In this study, mice were derived genetically deficient (null) in functional TGFα expression and carrying the Tg.AC/v-Ha-ras transgene. The goals were to determine if (a) papillomagenesis was dependent on TGFα and (b) progression to malignancy was dependent on TGFα expression. Groups of male and female mice heterozygous or homozygous for the TGFα null allele and hemizygous for the Tg.AC transgene were treated twice weekly for 10 or 15 wk with doses of 12-O-tetradecanoylphorbol-13-acetate (TPA) known to produce papillomas in Tg.AC mice. Papillomas were readily induced in both male and female TGFα null mice. Malignant progression of papillomas was observed in all TGFα null treatment groups. Additionally, we examined the response of TGFα null mice to full thickness dorsal wounds, a stimulus known to promote papillomagenesis in Tg.AC mice. As in the TPA study, papillomas were induced in both male and female TGFα null mice. These studies indicate that TGFα is not required for the induction and maintenance of papillomas nor is it essential for the malignant conversion of papillomas in Tg.AC mice.
1. Chomczynski P (1993). A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. BioTechniques. 15: 532–537.
2.
2. Chomczynski P and Sacchi N (1987). Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem.. 162: 156–159.
3.
3. Cook PW, Pittelkow MR, Keeble WW, Graves-Deal R, Coffey RJ Jr, and Shipley GD (1992). Amphiregulin messenger RNA is elevated in psoriatic epidermis and gastrointestinal carcinomas. Cancer Res.. 52: 3224–3227.
4.
4. Cox DR (1972). Regression models and life-tables. J. R. Stat. Soc.B34: 187–220.
5.
5. Derynck R (1986). Transforming growth factor-α: Structure and biological activities. J. Cell. Biochem.. 32: 293–304.
7. Derynck R, Goeddel DV, Ullrich A, Gutterman JU, Williams RD, Bringman TS, and Berger WH (1987). Synthesis of messenger RNAs for transforming growth factors α and β and the epidermal growth factor receptor by human tumors. Cancer Res.. 47: 707–712.
8.
8. Derynck R, Roberts AB, Winkler ME, Chen EY, and Goeddel DV (1984). Human transforming growth factor-α: Precursor structure and expression in E. coli. Cell38: 287–297.
10. Dlugosz AA, Cheng C, Williams EK, Darwiche N, Dempsey PJ, Mann B, Dunn AR, Coffey RJ Jr, and Yuspa SH (1995). Autocrine transforming growth factor α is dispensible for v-rasH3-induced epidermal neoplasia: Potential involvement of alternative epidermal growth factor receptor ligands. Cancer Res.. 55: 1883–1893.
11.
11. Dlugosz AA, Hansen L, Cheng C, Alexander N, Denning MF, Threadgill DW, Magnuson T, Coffey RJ Jr, and Yuspa SH (1997). Targeted disruption of the epidermal growth factor receptor impairs growth of squamous papillomas expressing the v-rasH3 oncogene but does not block in vitro keratinocyte responses to oncogenic ras. Cancer Res.. 57: 3180–3188.
12.
12. Finzi E, Kilkenny A, Strickland JE, Balaschak M, Bringman T, Derynck R, Aaronson S, and Yuspa SH (1988). TGFα stimulates growth of skin papillomas by autocrine and paracrine mechanisms but does not cause neoplastic progression. Mol. Carcinog.. 1: 7–12.
13.
13. Furstenberger G, Rogers M, Schnapke R, Bauer G, Hofler P, and Marks F (1989). Stimulatory role of transforming growth factors in multistage skin carcinogenesis: Possible explanation for the tumor-inducing effect of wounding in initiated NMRI mouse skin. Int. J. Cancer. 43: 915–921.
14.
14. Hansen LA and Tennant RW (1994). Focal transgene expression associated with papilloma development in v-Ha-ras-transgenic Tg.AC mice. Mol. Carcinog.. 9: 143–156.
15.
15. Higashiyama S, Abraham JA, Miller J, Fiddes JC, and Klagsburn M (1991). A heparin-binding growth factor secreted by macro-phage-like cells that is related to EGF. Science. 251: 936–939.
16.
16. Imamoto A, Beltran LM, and DiGiovanni J (1991). Evidence for autocrine/paracrine growth stimulation by transforming growth factor-α during the process of skin tumor promotion. Mol. Carcinog.. 4: 52–60.
17.
17. Jhappan C, Stahle C, Harkins RH, Fausto N, Smith GH, and Mer-lino GT (1990). TGFα overexpression in transgenic mice induces liver neoplasia and abnormal development of the mammary gland and pancreas. Cell. 61: 1137–1146.
18.
18. Jhappan C, Takayama H, Dickerson RB, and Merlino G (1994). Transgenic mice provide genetic evidence that transforming growth factor alpha promotes skin tumorigenesis via H-ras-dependent and H-ras-independent pathways. Cell. Growth Differ.. 5: 385–394.
19.
19. Jonckheere AR (1954). A distribution-free k-sample test against ordered alternatives. Biometrika. 41: 133–145.
20.
20. Kiguchi K, Beltran L, Dubowski A, and DiGiovanni J (1997). Analysis of the ability of 12-O-tetradecanoylphorbol-13-acetate to induce epidermal hyperplasia, transforming growth factor-α, and skin tumor promotion in wa-1 mice. J. Invest. Dermatol.. 108: 784–791.
21.
21. Kiguchi K, Beltran L, You J, Rho O, and DiGiovanni J (1995). Elevation of transforming growth factor-α mRNA and protein expression by diverse tumor promoters in SENCAR mouse epidermis. Mol. Carcinog.. 12: 225–235.
22.
22. Leder A, Kuo A, Cardiff RD, Sinn E, and Leder P (1990). v-Ha-ras transgene abrogates the initiation step in mouse skin tumori-genesis: Effects of phorbol esters and retinoic acid. Proc. Natl. Acad Sci. USA. 87: 9178–9182.
23.
23. Luetteke NC, Qiu T H, Peiffer RL, Oliver P, Smithies O, and Lee DC (1993). TGFα deficiency results in hair follicle and eye abnormalities in targeted and Waved-1 mice. Cell. 73: 263–278.
24.
24. Mann GB, Fowler KJ, Gabriel A, Nice EC, Williams RL, and Dunn AR (1993). Mice with a null mutation of the TGFα gene have abnormal skin architecture, wavy hair, and curly whiskers and often develop corneal inflammation. Cell. 73: 249–261.
25.
25. Marikovsky M, Breuing K, Liu PY, Eriksson E, Higashiyama S, Farber P, Abraham J, and Klagsbrun M (1993). Appearance of hep-arin-binding EGF-like growth factor in wound fluid as a response to injury. Proc. Natl. Acad. Sci. USA. 90: 3889–3893.
26.
26. Marquardt H, Hunkapiller MW, Hood LE, and Todaro GJ (1984). Rat transforming growth factor type 1 structure and relation to epidermal growth factor. Science. 223: 1079–1082.
28. Massague' J (1990). Transforming growth factor-α: A model for membrane-anchored growth factors. J. Biol. Chem.. 265: 21303–21396.
29.
29. Matsui Y, Halter SA, Holt JT, Hogan BLM, and Coffey RJ (1990). Development of mammary hyperplasia and neoplasia in MMTV-TGFα transgenic mice. Cell. 61: 1147–1155.
30.
30. Rho O, Beltran LM, Gimenez-Conti IB, and DiGiovanni D (1994). Altered expression of the epidermal growth factor receptor and transforming growth factor-α during multistage skin carcinogenesis in SENCAR mice. Mol. Carcinog.. 11: 19–28.
31.
31. Salomon DS, Kim N, Saeki T, and Ciardiello F (1990). Transforming growth factor α: An oncodevelopmental growth factor. Cancer Cells. 2: 389–397.
32.
32. Sandgren EP, Luetteke NC, Palmiter RD, Brinster RL, and Lee DC (1990). Overexpression of TGFα in transgenic mice: Induction of epithelial hyperplasia, pancreatic metaplasia, and carcinoma of the breast. Cell. 61: 1121–1135.
33.
33. Sandgren EP, Luetteke NC, Qiu TH, Palmiter RD, Brinster RL, and Lee DC (1993). Transforming growth factor alpha dramatically enhances oncogene-induced carcinogenesis in transgenic mouse pancreas and liver. Mol. Cell. Biol.. 13: 320–330.
34.
34. Shing Y, Christofori G, Hanahan D, Ono Y, Sasada R, Igaradhi K, and Folkman J (1993). Betacellulin: A mitogen from pancreatic β cell tumors. Science. 259: 1604–1607.
35.
35. Shoyab M, Plowman GD, McDonald VL, Bradley JG, and Todaro GJ (1989). Structure and function of human amphiregulin: A member of the epidermal growth factor family. Science. 243: 1074–1076.
36.
36. Siegel S (1956). Nonparametric Statistics for the Behavioral Sciences.McGraw-Hill, New York, pp. 116–127.
37.
37. Spalding JW, Momma J, Elwell MR, and Tennant RW (1993). Chemically induced skin carcinogenesis in a transgenic mouse line (Tg.AC) carrying a v-Ha-ras gene. Carcinogenesis. 14: 1335–1341.
38.
38. Tarone RE (1975). Tests for trend in life table analysis. Biometrika. 62: 679–682.
39.
39. Todaro GJ, De Larco JE, and Cohen S (1976). Transformation by murine and feline sarcoma viruses specifically blocks binding of epidermal growth factor to cells. Nature. 264: 26–31.
40.
40. Todaro GJ, Fryling C, and De Larco JE (1980). Transforming growth factors produced by certain tumor cells: Polypeptides that interact with epidermal growth factor receptors. Proc. Natl. Acad. Sci. USA. 77: 5258–5262.
41.
41. Vassar R and Fuchs E (1991). Transgenic mice provide new insights into the role of TGF-α during epidermal development and differentiation. Genes Dev.. 5: 714–727.
42.
42. Vaughn TJ, Paschall JC, and Brown KD (1992). Tissue distribution of mRNA for heparin-binding epidermal growth factor. Biochem. J.. 287: 681–684.
43.
43. Watanabe T, Shintani A, Nakata M, Shing Y, Folkman J, Igarashi K, and Sasada R (1994). Recombinant human betacellulin. J. Biol. Chem.. 269: 9966–9973.
44.
44. Wong ST, Winchell LF, McCune BK, Earp HS, Teixido' J, Massague' J, Herman B, and Lee DC (1989). The TGF-α precursor expressed on the cell surface binds to the EGF receptor on adjacent cells, leading to signal transduction. Cell. 56: 495–506.
