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A polymorphism in a let-7 microRNA binding site of KRAS in women with endometriosis.

Grechukhina O, Petracco R, Popkhadze S, Massasa E, Paranjape T, Chan E, Flores I, Weidhaas JB, Taylor HS - EMBO Mol Med (2012)

Bottom Line: Endometriosis is found in 5-15% of women of reproductive age and is more frequent in relatives of women with the disease.Activation of KRAS results in de novo endometriosis in mice, however, activating KRAS mutations have not been identified in women.We screened 150 women with endometriosis for a polymorphism in a let-7 microRNA (miRNA) binding site in the 3'-UTR of KRAS and detected a KRAS variant allele in 31% of women with endometriosis as opposed to 5% of a large diverse control population.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, USA.

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KRAS expression in hESCs obtained from endometrium of 10 women without endometriosis, eutopic endometrium of 10 women with endometriosis carrying WT KRAS and 10 women with endometriosis carrying the variant allele of KRAS gene at the LCS6 siteq-RT-PCR results show comparatively low levels of KRAS mRNA in normal endometrium, increased KRAS mRNA in hESC from women with endometriosis and the WT KRAS allele (p = 0.0007) and highest expression of KRAS mRNA in hESC carrying the variant KRAS allele (p = 0.00018 when compared to normal hESC, p = 0.0049 when compared to hESC from endometriosis patients with WT allele). (*, difference is significant when compared to normal hESC; **, difference is significant when compared to hESC from women with endometriosis homozygous for the WT KRAS allele).Western blot results show a 2.8-fold increase in KRAS protein in hESC with the variant allele.
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fig01: KRAS expression in hESCs obtained from endometrium of 10 women without endometriosis, eutopic endometrium of 10 women with endometriosis carrying WT KRAS and 10 women with endometriosis carrying the variant allele of KRAS gene at the LCS6 siteq-RT-PCR results show comparatively low levels of KRAS mRNA in normal endometrium, increased KRAS mRNA in hESC from women with endometriosis and the WT KRAS allele (p = 0.0007) and highest expression of KRAS mRNA in hESC carrying the variant KRAS allele (p = 0.00018 when compared to normal hESC, p = 0.0049 when compared to hESC from endometriosis patients with WT allele). (*, difference is significant when compared to normal hESC; **, difference is significant when compared to hESC from women with endometriosis homozygous for the WT KRAS allele).Western blot results show a 2.8-fold increase in KRAS protein in hESC with the variant allele.

Mentions: Quantitative polymerase chain reaction (after reverse transcription) (RT-PCR) was used to determine the level of KRAS mRNA in cultured hESC from women without endometriosis, women with endometriosis heterozygous for the variant allele, and women homozygous for the wild-type (WT) allele (n = 10 subjects per group). hESC from women without endometriosis were found to express approximately threefold lower levels of KRAS mRNA compared to both hESC from women with endometriosis carrying WT KRAS (KRAS/Actin: 0.001 ± 0.0002 vs. 0.003 ± 0.0004; p = 0.0007) and 10-fold lower mRNA levels compared to those carrying the variant KRAS allele (0.01 ± 0.002; p = 0.0001). KRAS mRNA was approximately threefold higher in hESC of subjects with the variant KRAS LCS6 compared to hESC from subjects with the non-variant allele (p = 0.0049; Fig 1a). Western blot analysis showed that hESC from women with the KRAS SNP had a 2.8-fold increase in KRAS protein compared to endometrium carrying the non-variant allele (Fig 1b).


A polymorphism in a let-7 microRNA binding site of KRAS in women with endometriosis.

Grechukhina O, Petracco R, Popkhadze S, Massasa E, Paranjape T, Chan E, Flores I, Weidhaas JB, Taylor HS - EMBO Mol Med (2012)

KRAS expression in hESCs obtained from endometrium of 10 women without endometriosis, eutopic endometrium of 10 women with endometriosis carrying WT KRAS and 10 women with endometriosis carrying the variant allele of KRAS gene at the LCS6 siteq-RT-PCR results show comparatively low levels of KRAS mRNA in normal endometrium, increased KRAS mRNA in hESC from women with endometriosis and the WT KRAS allele (p = 0.0007) and highest expression of KRAS mRNA in hESC carrying the variant KRAS allele (p = 0.00018 when compared to normal hESC, p = 0.0049 when compared to hESC from endometriosis patients with WT allele). (*, difference is significant when compared to normal hESC; **, difference is significant when compared to hESC from women with endometriosis homozygous for the WT KRAS allele).Western blot results show a 2.8-fold increase in KRAS protein in hESC with the variant allele.
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fig01: KRAS expression in hESCs obtained from endometrium of 10 women without endometriosis, eutopic endometrium of 10 women with endometriosis carrying WT KRAS and 10 women with endometriosis carrying the variant allele of KRAS gene at the LCS6 siteq-RT-PCR results show comparatively low levels of KRAS mRNA in normal endometrium, increased KRAS mRNA in hESC from women with endometriosis and the WT KRAS allele (p = 0.0007) and highest expression of KRAS mRNA in hESC carrying the variant KRAS allele (p = 0.00018 when compared to normal hESC, p = 0.0049 when compared to hESC from endometriosis patients with WT allele). (*, difference is significant when compared to normal hESC; **, difference is significant when compared to hESC from women with endometriosis homozygous for the WT KRAS allele).Western blot results show a 2.8-fold increase in KRAS protein in hESC with the variant allele.
Mentions: Quantitative polymerase chain reaction (after reverse transcription) (RT-PCR) was used to determine the level of KRAS mRNA in cultured hESC from women without endometriosis, women with endometriosis heterozygous for the variant allele, and women homozygous for the wild-type (WT) allele (n = 10 subjects per group). hESC from women without endometriosis were found to express approximately threefold lower levels of KRAS mRNA compared to both hESC from women with endometriosis carrying WT KRAS (KRAS/Actin: 0.001 ± 0.0002 vs. 0.003 ± 0.0004; p = 0.0007) and 10-fold lower mRNA levels compared to those carrying the variant KRAS allele (0.01 ± 0.002; p = 0.0001). KRAS mRNA was approximately threefold higher in hESC of subjects with the variant KRAS LCS6 compared to hESC from subjects with the non-variant allele (p = 0.0049; Fig 1a). Western blot analysis showed that hESC from women with the KRAS SNP had a 2.8-fold increase in KRAS protein compared to endometrium carrying the non-variant allele (Fig 1b).

Bottom Line: Endometriosis is found in 5-15% of women of reproductive age and is more frequent in relatives of women with the disease.Activation of KRAS results in de novo endometriosis in mice, however, activating KRAS mutations have not been identified in women.We screened 150 women with endometriosis for a polymorphism in a let-7 microRNA (miRNA) binding site in the 3'-UTR of KRAS and detected a KRAS variant allele in 31% of women with endometriosis as opposed to 5% of a large diverse control population.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, USA.

Show MeSH
Related in: MedlinePlus