| [Background] The mortality of ovarian cancer is the highest among the gynecologic tumors. 70 percent of the patients ,when going to doctor, have been in an advanced stage and most of them died of cancer metastasis . Therefore, to explore the molecular mechanism of invasion and metastasis and gene therapy of tumor have become one of the hot topics in the study on tumors. CD147 belongs to immunoglobulin superfamily, which not only participates in the proinflammation, but also highly expresses in varible tumor tissues. It not only promotes the invasion and metastasis of tumors, but also involves in tumors vascularization. The previous study on ovarian cancer in our laboratory indicated that CD 147 played an important role on the invasion and metastasis of ovarian tumors. After ovarian cancer cells were closed by CD147-antisense RNA, their transmembrane ability and invasiveness declined greatly, which showed CD 147 might be a drug target of ovarian cancer.[ Purpose] The present study is aimed to clarify the regulation ofCD 147 expression on ovarian cancers and to develop a human ovarian cancer orthotopic implantation high metastatic nude mice model for next gene therapy .[ Methods ] l.With Matinspector and Framework tools,we analysed the functions and structures in molecular biologic information of Homo sapiens CD147 mRNA complete sequences. 2.With Laser Confocal Technique and semi-quantity RT-PCR ,we studied qualitatively and quantitively on the CD 147 expression in ovarian cancer cell lines 3-AO and HO-8910PM under the condition of re-oxygenation after acute hypoxia, to observe hypoxia and reoxygenation regulation on the CD 147 expression and to explore the stability of CD 147 gene expression.3.With semi-quantity RT-PCR, we detected the expression of CD 147 mRNA in 3-AO and HO-8910PM under the condition of chemical hypoxia induced by CoCl2, to explore the regulation mechanism of hypoxia on CD 147 expression.4.At the premise of no human intervention, an ovarian tumor line of high metastasis, well-grown HO-8910PM, was implanted into nude mice subcutaneously and grew into tumor.5.With microsurgical technique,the tumor mass was taken out and implanted into the ovarian capsules of other nude mice ,and new formed tumor mass was sequentially passaged into next nude mice.6.The resected tumor tissue of every generation was observed with naked eyes, HE staining and Transmission Electronic microscope to verify the formation of tumor and the result of its metastasis.7.The genetic stability of xenografts was observed through chromosome analysis on the incubated cells and implanted tumor tissue.[Results] 1.103 high conservation matches with other species was found in Homo sapiens CD 147 mRNA complete sequences . In 3' enhancer of CD147mRNA ,there are two HIFF/HIF transcription binding sites which high conservation core sequences are similar to typical necleotide sequences of hypoxia response elements. 2.CD147 protein was located in cytoplasm. In 3-AO and HO-8910PM cell lines after acute hypoxia for 30 minutes, the CD 147 expression were remarkably higher compared with that in control group, and declined gradually to normal level after reoxygenation from 4 hours to 8 hours (P=0.000). Multiplex mean comparison by LSD-t: For HO-8910PM groups, no obvious difference existed between control and reoxygenation of 4 hours groups(p=0.557), while statistic significance was observed between the other groups(P < 0.05).For 3-AO groups, there was significant difference between all the groups(p < 0.05). 3.The expression of CD 147 mRNA increased gradually in a time-dependent manner with the elongation of hypoxia(r=0.998, p=0.338). At 24h after hypoxia the expression of CD 147 mRNA was still high. 4.With the elongation of reoxygenation, the expression of CD147mRNA droped in a time-dependent manner(r=0.904, P=0.035). 5. Tumors were formed in the ovaries of all the nude mice and metastasis occurred in three quarters of orthotopic implantation tumor. The natural survival time was 14-30 days on an average of 20.7 + 4.89 days. The m...
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