The Expression Of HMGI-C In Uterine Leiomyomas Cells And The Influence Of Anti Sense Oligodeoxy Nucleotide With Uterine Leiomyomas Cells Proliferation

Uterine leiomyomata (UL) is associated with a variety of characteristic cytogenetic abnormalities. The significance of these chromosomal aberrations in the pathobiology of UL remains to be determined . UL is one of several benign tumors characterized by frequent chromosomal rearrangement involving 12q15. The 12q15 rearrangement in leiomyomata typically is manifested as t(12;14)(q15;q23-24) which has been hypothesized to create pathobiologically significant fusion transcript and influence in the progression of UL. HMGI-C belongs to the HMG (high mobility group) protein family .The HMG family of protein are small non histone nuclear proteins which play an important role in the regulation of chromatin structure and function as a architectural factor to mediate structural changes in DNA. The HMGI-C gene was localized to chromosome 12 band q15 region often rearranges in benign mesenchymal tumors. However HMGI-C dysregulation as a result of specific rearrangement involving 12q15, the chromosomal site in which the HMGI-C gene was localized was also identified in UL. Considerable interest has recently been shown in HMGI-C, its expression is involve in neoplastic tumors and and apparently in tumorigenesis, whereas no expression could be found in normal tissue adjacent to the tumor. Although some evidence suggest a role of HMGI-C in regulation of cell proliferation, the precise function of HMGI-C in the development or progression of UL remains unclear and to be determined. In this study ,we would explore the role and possible mechanism of the HMGI-C protein in uterine leiomyomatas. A total of 102 cases UL specimens and the normal myometrium adjacent to the UL, were obtained from 86 patients undergoing myomectomy or hysterectomy and a portion of each specimens for one-step reverse transcriptase polymerase chain reaction (RT-PCR) analyze. HMGI-C gene mRNA were detected in experiment from 56 cases out of 102 (54.90%) UL and no one out of 102 cases of normal myometrium . On the basis of these results, the specimens were categorized as HMGI-C positive UL group (Group Ⅰ) and HMGI-C negative UL group(GroupⅡ), normal myometrium group (GroupⅢ) . We also detected the expression of HMGI-C gene proteins in three experimental groups using immunohistochemistry analysis and the results suggested that there was over-expression of HMGI-C gene in UL than normal comparison groups. It was indicated that HMGI-C could be a diagnostic marker for UL. It may have a possible role for the proliferation of myometrium cells. The present study investigated that there was relationship between HMGI-C over expression and the size of UL.The maximum diameter of each UL was measured and the Mean UL diameter was 7.8±0.24 cm with a range of 1.0～35.2 cm . The mean UL diameter among specimens of UL with expression group of HMGI-C gene was significantly greater than UL with non-expression group of HMGI-C(13.2±2.9 cm versus 6.8±3.2 cm ; p<0.001). The diameter of UL >7.8 cm demonstrated a significantly higher proportion of HMGI-C positive expression when compared with UL of diameter <7.8 cm (83.8% versus 40.0%, p<0.01). In summary, there was a significant relationship existing between expression of HMGI-C gene and UL size, and suggested that the expression of HMGI-C gene might enhance the cells growth of UL. These results indicated that the HMGI-C played a critical role in the growth and differentiation of UL cells and provided the first insights that HMGI-C over-expression should be evaluated as a potential diagnosis and prognostic marker in UL. 20 cases were taken from groupⅠand groupⅡ,10 cases from groupⅢrespectively and discretionarily. All of their chromosomal karyotype of UL were detected and analyzed. There were 5 cases (5/20, 25%) of abnormal karyotypes with t(12;14) (q15;q23～24) in groupⅠ,but no abnormal karyotype found in other groups was 5 cases. Depend on this result the group Ⅰwas divided into groupⅠa andⅠb. GroupⅠa was the UL with chromosome rearrangement involving t(12;14)(q15,q23-24) and groupⅠb with normal karyotype. Detecting the expression of HMGI-C gene in group Ⅰa andⅠb respectively by immunohistochemistry analysis, it showed that the level LI(Labeling index) of HMGI-C gene in groupⅠa was signifcantlyhigher than groupⅠb. It suggested that the chromosome rearrangement involving t12q15 increased the expression of HMGI-C gene in the tissues of UL and it was also related to the occurrence and growth of uterine myoma. There are two members in estrogen receptor(ER), ERαand ERβ. ERαwas localized on the chromosome 6 band q25.1 and ERβwas localized on the chromosome 14 band q23～24. The latter region often have been rearranged in UL, for example t(12;14)(q15;q23～24). Just as above said, HMGI-C which was localized on the chromosome 12 band q15.Therefor, ER βhave a possibility to be a transcriptional candidate partner of HMGI-C in t(12;14). In present study we also detected the protein expression of ERβand PCNA gene in groups Ⅰ,Ⅱ,Ⅲ,ⅣandⅠa ,Ⅰb. The results showed that the levels of protein expression of ERβand PCNA gene in every groups have the same trend of the HMGI-C gene. It suggested that HMGI-C played a critical role in proliferation of UL cells through increasing or cooperating with ERβgene expression. The mechanism of this function of HMGI-C remains to be researched. In the last part, we evaluated the effect of anti sense oligodeoxy nucleotides (ASODN) of HMGI-C on human uterine leiomyoma cell growth. The UL cell lines(ULCL) which were identified by α-actin antibody and could be steadily generated were constructed for the first time. Through transfecting the different concentration of HMGI-C of ASODN into ULCL cell by the liposome method. It illuminated the dialysis of ASODN and the function of repression on proliferation...