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  • Title: [Construction and expression of recombinant cecropin B-binding site of luteinizing hormone releasing hormone gene and its anticancer function].
    Author: Li XY, Li HL, Zheng GY.
    Journal: Zhonghua Fu Chan Ke Za Zhi; 2007 Jul; 42(7):477-81. PubMed ID: 17961339.
    Abstract:
    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. CONCLUSION: 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.
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