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ORIGINAL RESEARCH ARTICLE

Growth Repression in Diethylstilbestrol/Dimethylbenz[a]anthracene–Induced Rat Mammary Gland Tumor Using Hecate-CGß Conjugate

Monika Zaleska^*, Agnieszka Waclawik^*, Gabriel Bodek^*, Anna Zezula-Szpyra^*, Xiangdong Li, Tomasz Janowski, Wiliam H. Hansel, Nafis A. Rahman and Adam J. Ziecik^*,1

^* Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, and Department of Obstetrics and Pathology of Reproduction, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, 10-747 Olsztyn, Poland; Pennington Biomedical Research Center, Louisiana State University, Baton Rouge, Louisiana 70808-4124; and Department of Physiology, University of Turku, 20520 Turku, Finland

To whom requests for reprints should be addressed at ¹ Institute of Animal Reproduction and Food Research Polish Academy of Sciences, Tuwima 10, 10-747 Olsztyn, Poland. E-mail: ziecik{at}pan.olsztyn.pl

Recently, we have shown that Hecate-CGß conjugate, which is a fusion of the lytic peptide Hecate and a 15–amino acid fragment of the ß-chain of chorionic gonadotropin (CGß), selectively destroys mammary gland carcinoma cells that possess luteinizing hormone receptors (LHR) in vitro. We induced mammary gland tumors using combined prenatal exposure to synthetic diethylstilbestrol (DES) and additional postnatal exposure to dimethylbenz[a]anthracene (DMBA). Rats with tumors were equally randomized (10/group) and treated with either sham (control) or 12 mg/kg body wt of either Hecate or Hecate-CGß once a week for 3 weeks by tail vein injections. One week after the last injection, rats were killed. Reverse-transcription–nested polymerase chain reaction/Southern blotting revealed alternatively spliced mRNA for LHR in tumor tissues of 5 of 30 females, which was further confirmed by Western blot analysis. The percentage of tumor volume increase was lowest in the group treated with Hecate-CGß (45.3 ± 27.6), compared with Hecate- and sham-treated, control group (324.8 ± 78.1 and 309.9 ± 51.2, respectively; P < 0.001). Hecate-CGß induced a significant decrease in tumor burden compared with controls (9.5 ± 2.1 mg/g body wt vs. 21.6 ± 2.9; P < 0.01). A smaller reduction in tumor burden was also observed in Hecate-treated females (17.6 ± 1.6 mg/g body wt vs. 21.6 ± 2.9; P < 0.05). Our results prove the principle that Hecate-CGß conjugate is able to repress mammary gland tumor growth, even in tumor tissues that lack or have very low levels of LHR. The mechanism of Hecate-CGß conjugate action in repression of DES/DMBA-induced tumor growth needs to be further analyzed to clarify the molecular mechanisms of Hecate-CGß conjugate action in vivo.

Key Words: mammary gland tumor • LH receptor • Hecate-CGß conjugate • cancer treatment

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