G蛋白偶联受体30和磷酸化AKT在子宫内膜腺癌中表达及意义

目的:探讨G蛋白偶联受体30(GPR30)和磷酸化AKT(P-AKT)在子宫内膜腺癌发生发展中的作用及相互关系。方法:用免疫组化SP法检测10例增生期子宫内膜,49例子宫内膜增殖症,55例子宫内膜腺癌组织中GPR30和P-AKT的表达。结果:GPR30蛋白在子宫内膜腺癌、子宫内膜增殖症的阳性表达率(81.8%、67.3%)均明显高于增生期子宫内膜中的阳性表达率(20.0%);子宫内膜增殖症中GPR30蛋白在单纯型增生子宫内膜(SH)、复杂型增生子宫内膜(CH)和不典型增生子宫内膜(AH)的阳性表达率分别为30.0%(3/10),70.0%(14/20)和84.2%(16/19),AH和SH的差异有统计学意义(P=0.012)。P-AKT在子宫内膜腺癌、子宫内膜增殖症的阳性表达率(78.2%、71.4%)均显著高于增生期子宫内膜中的阳性表达率(20.0%);子宫内膜增殖症中P-AKT蛋白在SH、CH、AH的阳性表达率分别为40.0%(4/10),65.0%(13/20)和94.7%(18/19),AH与SH的差异有统计学意义(P=0.005)。子宫内膜腺癌GPR30、P-AKT蛋白阳性表达率与组织分化程度、FIGO分期及患者是否绝经有关,在中、低分化组(P=0.023;P=0.009)、FIGOⅡ～Ⅲ期(P=0.039;P=0.017)及绝经后(P=0.046;P=0.031)较高。肌层浸润较深的子宫内膜腺癌P-AKT蛋白表达水平高于肌层浸润较浅者(P=0.042)。子宫内膜腺癌组织中GPR30与P-AKT蛋白表达呈正相关(P<0.001)。结论:GPR30和P-AKT活化与子宫内膜癌的发生发展有关。     

G蛋白偶联雌激素受体在子宫内膜腺癌组织表达及其意义

目的探讨G蛋白偶联雌激素受体(GPER)在子宫内膜腺癌组织中发生发展的作用。方法选取2005年3月至2008年4月郑州大学第一附属医院病理科手术及活检标本的存档蜡块,采用免疫组织化学SP法检测55例子宫内膜腺癌、49例子宫内膜增殖症及10例正常增生期子宫内膜组织中GPER蛋白的表达,分析其与子宫内膜腺癌患者临床病理特征的关系。结果正常增生期子宫内膜组织、子宫内膜增殖症组织和子宫内膜腺癌组织中GPER蛋白的阳性表达率分别为20.0%、67.3%和81.8%,差异具有统计学意义(χ2=15.778,P<0.001)。子宫内膜增殖症中,简单型增生、复杂型增生及不典型增生内膜组织中GPER蛋白的阳性表达率分别为30.0%、70.0%和84.2%,差异具有统计学意义(χ2=8.864,P=0.012)。绝经后和未绝经子宫内膜腺癌组织中GPER的阳性表达率分别为89.7%和62.5%,差异具有统计学意义(χ2=3.977,P=0.046)。高分化和中、低分化子宫内膜腺癌组织中GPER的阳性表达率分别为71.4%和100%,差异具有统计学意义(χ2=5.196,P=0.023)。临床分期Ⅰ期和(Ⅱ+Ⅲ)期子宫内膜腺癌组织中GPER的阳性表达率分别为73.0%和100%,差异具有统计学意义(χ2=4.268,P=0.039)。有淋巴结转移和无淋巴结转移的子宫内膜腺癌组织中GPER的阳性表达率分别为66.7%和83.7%,差异无统计学意义(χ2=0.210,P=0.646)。在肌层浸润深度<1/2和≥1/2的子宫内膜腺癌组织中GPER的阳性表达率分别为75%和100%,差异无统计学意义(χ2=3.057,P=0.080)。结论 GPER与子宫内膜腺癌的发生、发展可能具有相关性。     

G protein-coupled estrogen receptor 1 (GPER 1) mediates estrogen-induced,proliferation of leiomyoma cells

Abstract

G protein-coupled estrogen receptor 1 (GPER-1, formerly known as GPR30) has been proposed as the receptor for estrogen-induced, growth of leiomyomas though its precise mechanisms of action are not clear. We obtained leiomyoma cells (LC) and normal smooth muscle cells from 28 women (n?=?28, median age 38 years, median parity 1.0). We incubated them with 17-β estradiol (E₂), after blocking, or upregulating, expression of GPER-1 with ICI182,780 (a GPER-1 agonist) and siGPR30, respectively. We evaluated the role of GPER-1 in the mitogen-activated protein kinase (MAPK) signaling pathway using Western blot analysis. We studied cell proliferation with 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide, and, mitotic activity with phosphohistone H3 (PPH3) expression in leiomyoma, and, matched, normal, smooth muscle tissues using standard immunohistochemistry. Downregulation of GPER-1 expression with siGPR30 partially attenuated the E₂-activated MAPK signaling pathway (p?<?0.01). Upregulation of GPER-1 with ICI182,780 enhanced the E₂-activated MAPK signaling pathway (p?<?0.01). ICI182,780 enhanced E₂-induced proliferation of LC (p?<?0.01), while knock down of the GPER-1 gene with GPER-1 small interfering RNA partially inhibited E₂-induced cell proliferation (p?<?0.01). There were no significant differences in PPH3 expression between LCs and normal smooth muscle tissues (p?>?0.05). Neither ICI182,780 nor siGPR30 increased mitosis in LCs (p?>?0.05). Our results indicate that GPER-1 mediates proliferation of estrogen-induced, LC by activating the MAPK pathway, and, not by promoting mitosis.     

GPR30在宫颈癌细胞生长中的作用及其机制研究

目的:体外研究雌激素膜受体GPR30对宫颈癌细胞生长的影响及其作用机制。方法:选择宫颈腺癌HeLa和宫颈鳞癌SiHa细胞株,分别用GPR30特异性激动剂G1和拮抗剂G15处理宫颈癌细胞株。RT-PCR、Western blot法检测处理前后宫颈癌HeLa与SiHa细胞中GPR30、TLR3 mRNA及其蛋白表达变化;MTT法检测G1、G15及Poly I:C处理对宫颈癌细胞生长的影响。结果:(1)HeLa细胞中GPR30表达量高于SiHa细胞。G1处理后HeLa、SiHa细胞中GPR30 mRNA及其蛋白表达水平增高,与对照组比较,差异有统计学意义(P0.05;P0.05);G15处理后,HeLa、SiHa细胞中GPR30 mRNA及其蛋白表达水平降低,与对照组比较,差异有统计学意义(P0.05;P0.05)。(2)HeLa、SiHa细胞中TLR3 mRNA表达量分别为(0.5327±0.05373)、(0.3526±0.05774),蛋白表达量分别为(0.3572±0.097039)、(0.5002±0.09718)。G1能降低He La、Si Ha细胞中TLR3mRNA及其蛋白表达水平,G1 10-6mol/L处理组与对照组比较差异有统计学意义(P均0.05)。G15能增高HeLa、SiHa细胞中TLR3 mRNA及其蛋白表达水平,G15 10-5mol/L处理组与对照组比较,差异有统计学意义(P均0.05),与Poly I:C处理组比较差异无统计学意义(P0.05)。(3)10-6mmol/L、10-5mmol/L G1分别处理后,宫颈癌HeLa、SiHa细胞生长增殖率分别为(16.68±5.86)%、(26.67±3.25)%及(14.99±6.43)%、(22.72±1.77)%,与空白对照组相比,差异有统计学意义(P均0.05)。10-6mmol/L、10-5mmol/L G15分别处理后,宫颈癌HeLa、SiHa细胞的生长抑制率分别为(21.09±2.32)%、(22.99±3.15)%及(15.86±6.49)%、(19.18±2.61)%,与空白对照组相比,差异有统计学意义(P均0.05)。结论:宫颈癌细胞中存在雌激素膜受体GPR30表达,宫颈腺癌细胞中GPR30表达量高于宫颈鳞癌细胞,体外调节GPR30表达可影响宫颈癌细胞生长。抑制GPR30表达可通过上调TLR3表达而抑制宫颈癌细胞生长,GPR30可能成为宫颈癌治疗的新靶点。     

G蛋白耦联受体30(GPR30)在上皮性卵巢癌中的表达与增殖相关基因c-fos、cyclinD1表达的相关性

目的:探索新型雌激素受体G蛋白耦联受体30(G protein coupled receptor 30,GPR30)在人上皮性卵巢癌发生发展中的作用。方法:采用免疫组织化学SP法检测30例上皮性卵巢癌组织中GPR30、增殖相关基因c-fos和cyclinD1的表达,并以9例良性卵巢肿瘤、4例正常卵巢组织作对照。结果:GPR30在上皮性卵巢癌的表达水平(80.0%)显著高于良性卵巢肿瘤(44.4%)和正常卵巢组织(25.0%)(P<0.01;P<0.05)。GPR30和c-fos的表达与上皮性卵巢癌的病理类型、FIGO分期有关,在浆液性囊腺癌、FIGOⅢ-Ⅳ期的表达水平显著升高(P<0.05)。未见cyclinD1的表达与上皮性卵巢癌临床病理参数的关系(P>0.05)。上皮性卵巢癌中GPR30与c-fos、cyclinD1的表达具有正相关性(P<0.01;P<0.05)。结论:GPR30可能通过c-fos、cyclinD1促进卵巢癌增殖,参与上皮性卵巢癌的发生、发展。     

Association between genetic polymorphisms in androgen receptor gene and the risk of preeclampsia in Korean women

Purpose  

To investigate associations between the androgen receptor (AR) polymorphisms as CAG repeats, GGC repeats and c.211G>A polymorphism and the risk of preeclampsia.     

临产前后子宫平滑肌组织中G蛋白偶联受体30的表达及其临床意义研究

目的 探讨G蛋白偶联受体30（GPR30） mRNA和蛋白在临产前、后子宫平滑肌组织中的表达及其与分娩发动的相关性。方法 2011年3月至2012年3月在青岛市市立医院选取临产前进行选择性剖宫产的足月妊娠妇女作为未临产组（n=40）,自然临产进入产程活跃期后进行急症剖宫产的足月妊娠妇女作为临产组（n=50）,于剖宫产术中取少量子宫平滑肌组织,采用实时荧光定量PCR技术和Western blot技术检测两组子宫平滑肌组织中GPR30 mRNA和蛋白的表达量;并于术前取静脉血,采用放射免疫法测定血清中雌二醇（E2）、雌三醇（E3）以及孕酮（P）的含量。结果 实时荧光定量PCR方法检测临产组和未临产组子宫平滑肌组织中均有GPR30 mRNA表达,临产组样本△Ct为3.08±0.35,未临产组样本△Ct为4.02±0.67,差异具有统计学意义（P＜0.05）,用双△Ct法进行相对定量分析,临产组GPR30 mRNA表达量是未临产组的1.92倍。Western blot技术检测临产组和未临产组子宫平滑肌组织中均有GPR30蛋白表达,临产组GPR30蛋白相对表达量4.06±0.25明显高于未临产组1.94±0.23,差异有统计学意义(P＜0.05)。临产组血清E2、E3浓度[（7882.41±921.76）pmol/L,（544.12±15.58）nmol/L]均显著高于未临产组[（5210.72±873.13）pmol/L,（326.36±12.26）nmol/L]。结论 足月妊娠分娩发动时GPR30高表达可能参与介导雌激素对子宫收缩的调节作用。     

Leptin receptor gene polymorphisms in severely pre-eclamptic women

Variants of the leptin receptor gene (LEPR) may modulate the effect of elevated serum leptin levels in pre-eclampsia. The aim of our study was to evaluate the LEPR gene polymorphisms Lys109Arg (A109G) and Gln223Arg (A223G) in severely pre-eclamptic women. In a case–control study, we analyzed blood samples from 124 severely pre-eclamptic patients and 107 healthy control women by the polymerase chain reaction–restriction fragment length polymorphism method. The Pearson χ² test was used to estimate odds ratios (OR) and 95% confidence intervals (CI). The association was adjusted for maternal age, pre-pregnancy body mass index and primiparity with logistic regression analysis. Pregnant women with the LEPR 223G allele (223A/G or 223G/G genotype) had almost double the risk of developing severe pre-eclampsia compared with patients with the 223A/A genotype (adjusted OR = 1.92, 95% CI: 1.07–3.41). Genotype variants of LEPR A109G alone did not affect the risk of severe pre-eclampsia. Haplotype estimation of A109G and A223G polymorphisms of the LEPR gene revealed that the G-A haplotype versus other pooled haplotypes was significantly less common in the pre-eclamptic group (p < 0.01), while the G-G haplotype versus others was overrepresented among severely pre-eclamptic patients (p < 0.01), compared with controls. In conclusion, our data indicate that LEPR A223G polymorphism may individually modify the risk of severe pre-eclampsia.     

表皮生长因子受体在子宫腺肌病中的表达及其临床意义

目的：研究表皮生长因子受体（ＥＧＦＲ）在子宫腺肌病中的组织定位和表达；探讨ＥＧＦＲ表达与子宫腺肌病发生的关系，寻求经调控ＥＧＦ／ＥＧＦＲ治疗子宫腺疾病的新途径。方法：应用免疫组织化学ＡＢＣ法检测子宫内膜组织１０份和子宫腺肌病组织３０份标本的ＥＧＦＲ定位和表达以及月经周期中内膜组织的ＥＧＦＲ表达强度变化。结果：正常子宫内膜和子宫腺肌病病灶均存在ＥＧＦＲ表达；ＥＧＦＲ位于内膜腺上皮和间质及平滑肌细胞上；表达强度：腺体＞间质＞平滑肌；子宫内膜＞异位内膜，但无统计学差异；ＥＧＦＲ表达无周期性变化，异位内膜侵入程度与临床症状有相关性。结论：正常子宫内膜和子宫腺肌病异位内膜存在ＥＧＦＲ共同表达；提示两者具有组织同源性；异位内膜中ＥＧＦＲ表达可能与子宫腺肌病发生机制有关。经降调节ＥＧＦ／ＥＧＦＲ表达可望成为治疗子宫腺肌病的新途径。     

子宫腺肌病中孕激素受体B表达及甲基化的研究

目的:探讨子宫腺肌病中孕激素受体B(progesterone receptor B,PR-B)表达及其甲基化在疾病发生中的作用。方法:用免疫组化法测定PR-B的表达,甲基化特异性基因扩增(methylation-specific PCR,MSP)方法检测50例子宫腺肌病患者在位与异位内膜和18例输卵管不孕或良性卵巢囊肿患者子宫内膜中PR-B的甲基化状态,并分析甲基化与PR-B表达的相关性。结果:子宫腺肌病患者在位和异位内膜PR-B表达明显低于对照内膜(P0.01);子宫腺肌病异位内膜中PR-B甲基化比率显著高于对照内膜(P0.05);且子宫腺肌病在位和异位内膜中PR-B甲基化状态与PR-B表达相关(P0.05)。结论:PR-B低表达及其甲基化可能与子宫腺肌病的发生有关。     

The accuracy of transvaginal ultrasound and uterine artery Doppler in the prediction of adenomyosis

ObjectiveTo measure the accuracy of the ultrasonographic features in predicting adenomyosis and to determine if there is a role for uterine artery Doppler in adenomyosis prediction.Study designA prospective comparative study.SettingCairo University hospital.Materials and methodsThree hundred and fifty-two women who were scheduled for hysterectomy for various indications underwent preoperative transvaginal ultrasound scan (TVS) and uterine artery Doppler velocimetry in an attempt to diagnose adenomyosis. All the results were then correlated with histopathological results after hysterectomy.ResultsForty-eight participants were ultrasonographically diagnosed as having adenomyosis from which 37 patients were histologically confirmed. Both groups were comparable in age, but adenomyosis tend to occur in multiparas. We found that subendometrial linear striations, myometrial cysts’ number and poor endometrial delineation were significantly associated with adenomyosis. Sensitivity, specificity, positive predictive value, negative predictive value and overall accuracy of TVS for diagnosing adenomyosis were: 75.68%, 90.79%, 49.12%, 96.95% and 89.20%, respectively. Heterogenous myometrial echotexture was the most common ultrasonographic feature in adenomyotic cases. Neither uterine artery resistance index nor pulsatility index showed significant association with adenomyosis.ConclusionTVS is a potentially valuable tool in predicting adenomyosis especially when subendometrial linear echogenic striations, myometrial cysts, and poor endometrial delineations were found. However, uterine artery Doppler has no diagnostic values.