Human Papillomavirus E6 Knockdown Restores Adenovirus Mediated-estrogen Response Element Linked p53 Gene Transfer in HeLa Cells.

The p53 gene is inactivated by the human papillomavirus (HPV) E6 protein in the majority of cervical cancers. Treatment of HeLa S3 cells with siRNA for HPV E6 permitted adenovirus-mediated transduction of a p53 gene linked to an upstream estrogen response element (ERE). Our previous study in non-siRNA treated HHUA cells, which are derived from an endometrial cancer and express estrogen receptor ß, showed enhancing effects of an upstream ERE on adenovirus-mediated p53 gene transduction. In HeLa S3 cells treated with siRNA for HPV E6, adenovirus-mediated transduction was enhanced by an upstream ERE linked to a p53 gene carrying a proline variant at codon 72, but not for a p53 gene with arginine variant at codon 72. Expression levels of p53 mRNA and Coxsackie/adenovirus receptor (CAR) mRNA after adenovirus-mediated transfer of an ERE-linked p53 gene (proline variant at codon 72) were higher compared with those after non-ERE-linked p53 gene transfer in siRNA-treated HeLa S3 cells. Western blot analysis showed lower ß-tubulin levels and comparatively higher p53/ß-tubulin or CAR /ß-tubulin ratios in siRNA-treated HeLa S3 cells after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with those in non-siRNA-treated cells. Apoptosis, as measured by annexin V binding, was higher after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with that after non-ERE-linked p53 gene transfer in siRNA-treated cells.

An upstream estrogen response element linked to exogenous p53 tumor suppressor gene expression differentiates effects of the codon 72 polymorphism.

The objective of this study was to assess the effects of an upstream estrogen response element (ERE) on exogenous p53 tumor suppressor gene with a codon 72 polymorphism about which there have been controversial reports in relation to cancer risk. The p53 gene (bases 166-1143 from start codon) with the codon 72 polymorphism, inserted into the pIRES-hrGFP II plasmid with or without upstream ERE, were transfected into HHUA endometrial cancer cells expressing the estrogen receptor. The ERE-linked p53 gene with the proline variant at codon 72 showed lower transfection rates than the gene without ERE or with the arginine variant at codon 72. p21 expression was significantly higher in HHUA cells transfected with the proline variant gene than in those transfected with the arginine variant gene. We consider that the presence of an upstream ERE promotes the transcriptional effects of the exogenous p53 gene with the proline variant, which strengthens the expression of p21, and results in lower transfection rates through cell cycle inhibition.

Transcervical microwave myolysis for uterine myomas assisted by transvaginal ultrasonic guidance.

AIM: The effects of transcervical microwave myolysis at 2.45 GHz after microwave endometrial ablation for menorrhagia were examined in patients with myomas. METHODS: A transcervical microwave irradiation system assisted by transvaginal ultrasonic guidance was developed. Ten patients waiting for microwave endometrial ablation for menorrhagia caused by myomas entered the study after complete informed consent was obtained. Their outcomes were examined at least 6 months after transcervical microwave myolysis using the system. After completion of microwave endometrial ablation, interstitial microwave irradiation was performed using a guiding needle set in a puncture adaptor attached to a transvaginal ultrasonic probe and a microwave applicator of 1.6 mm in diameter. Shrinkage of the myoma was measured at 3 months and > or =6 months after the operation. RESULTS: In nine patients with typical myomas, the major part consisting of a submucous or an intramural node 4.0-7.5 cm in size was necrotized. The typical myomas had shrunk by 41-68% at 3 months and 37-69% at > or =6 months after the operation. In one patient with a 6.8-cm cellular leiomyoma, necrosis was limited to the neighborhood of the applicator tip. Shrinkage was 17% at both 3 and 6 months. This patient required a second microwave endometrial ablation at 6 months to treat recurrent menorrhagia. None of the patients underwent hysterectomy after the treatment. No remarkable complications were encountered. CONCLUSIONS: Typical myomas shrink after microwave myolysis following microwave endometrial ablation. Transcervical microwave myolysis seems to be applicable as a low-invasive treatment for a typical myoma.

Endometrial thickness in Japanese women with hypertension or/and type 2 diabetes mellitus.

OBJECTIVES: The purpose of this study was: (a) to examine whether the endometrium of postmenopausal women with hypertension (HT) and/or type 2 diabetes mellitus (DM) was thicker than that of healthy controls (HC) and (b) whether endometrial thickness (ET) was associated with endometrial cancer risk factors. STUDY DESIGN: A total of 242 postmenopausal women were included in this study. Thirty women with type 2 DM, 49 women with HT, 23 women with DM and HT and 140 HCs were studied. Clinical evaluation of all women was done using TVS and endometrial cytology. RESULTS: In the 140 HCs, the mean ET of nulliparous women was larger than multiparous women (3.0 mm versus 1.6 mm, p = 0.017). However, there was no significant relationship between ET and other clinical parameters such as a history of sterility or BMI. There was no significant difference in ET amongst the four groups. The relationship between ET, age, pregnancy history, menopause age, BMI and presence of DM or HT were analyzed using stepwise multivariate analysis. There was no significant relationship between these risk factors and ET (p = 0.063). CONCLUSIONS: The risk factors such as age, pregnancy history, menopause age, BMI and presence of DM or HT were not related to ET in Japanese women.