| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
The results previously cleared in this study are four points as follows ;
1.The hyperinethylation of hMLH1 promoter is found about 40% of endometrial cancers.
2.Protein expression is decreased with hyperinethylation of hMLH1 promoter.
3.The hypermethylation is already found in normal endoinetrium adjacent to endometrial cacers.
4.Genetic instability is also found both in cancer and normal endometrium with hypermethylation of hMLH1.
With the above-mentioned results, the followings are cleared in 2003.
1.Quantitative methylation analysis of hMLH1 promoter
Methylation specific PCR(MSP)was used as qualitative analysis, however, quantitative analysis for hMLH1 promoter hypermetylation is required.For this purpose, we used bisulfite sequencing to investigate metylation status within 700-bp promoter area of hMLH1.With the 56 endometrial cancer cases, 16 cases (29%) are fully methylated in promoter, 14 cases(25%) are partially methylated, and 14 cases (46%) are non-methylated.
2. Relation between hMLH1 methylation and PTEN mutation
It is reported that PTEN mutations are found in early stage of endometrial carcinogenesis.In this study, PTEN mutations are recognized in 40% of endometrial cancers, and in particular, insertion/deletion mutations of PTEN are significantly related with hMLH1 promoter hypermethylations.
3.Laser capture microdissection : LCM
LCM is used to distinguish carcinoma or precancerous lesion with normal stromal tissue.Methylation analysis was technically difficult with LCM because of the small amount of DNA, but we solved this problem by the combination of PCR and methylation sensitive restriction enzymes.As a result, methylation frequency of the endoinetrial hyperplasia and cancer was almost equal, While the methylation frequency of the normal endometrium was zero.
With these results, our future plans are elucidation of endometrial carcinogenesis, and clinical application of endometrial cancer screening.
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