[Construction and expression of recombinant cecropin B-binding site of luteinizing hormone releasing hormone gene and its anticancer function].
OBJECTIVE
To construct and express recombinant cecropin B-binding site of luteinizing hormone releasing hormone (CB-LHRH') gene, and to evaluate the anticancer function of CB-LHRH' on human ovarian cancer cell line SKOV3 and human endometrial cancer cell line HEC-1B.


METHODS
The sequence of the cDNA encoding CB-LHRH' was designed, artificially synthesized, verified by DNA sequence analysis and expressed by Bac-to-Bac baculovirus expression system. The expression of CB-LHRH' proteins were identified by western dot blot using rabbit polyclonal antibody against LHRH as the primary antibody. To determine the anticancer effects of the CB-LHRH' protein, ovarian cancer cell line SKOV3 and endometrial adenocarcinoma cell line HEC-1B were treated by different doses of the CB-LHRH' protein. Cell growth inhibition assay was performed using the 2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl) 5 [(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) kit at different times, and cell morphologic changes were observed under the inverted microscope.


RESULTS
The inhibitory rate of proliferation by CB-LHRH' increased with the increase of dose and time respectively: SKOV3 cell, from (5.03 +/- 0.08)% to (53.24 +/- 1.22)%; HEC-1B cell, from (5.13 +/- 0.37)% to (56.16 +/- 1.08)%. The inhibitory effect on HEC-1B cell was stronger than that on SKOV3 cell (P < 0.01). There were obvious cell morphologic changes.


CONCLUSIONS
The CB-LHRH' protein has anticancer activity, probably through the LHRH receptor, and it may be expected to be a potential candidate as peptide drugs for targeted cancer therapy.
Li
Xiao-yong
XY
Department of Gynecology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China.
Li
He-lian
HL
Zheng
Gui-ying
GY
D000818:Animals; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology*; D016367:Baculoviridae / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D016889:Endometrial Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D006801:Humans; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D018411:Spodoptera; D013997:Time Factors; D014162:Transfection
D018931:Antineoplastic Agents, Hormonal; D011994:Recombinant Proteins; D007987:Gonadotropin-Releasing Hormone