子宮頚癌の発生における分子機構解析,特にFHIT遺伝子の変異とHPV E6-E7遺伝子の発現相関について

取得学位 : 博士（医学）, 学位授与番号 : 医博乙第1572号, 学位授与年月日 : 平成15年2月5日, 学位授与大学 : 金沢大

子宮体癌に対するアロマターゼ遺伝子をターゲットとした遺伝子治療への戦略

金沢大学附属病院【目的】子宮体癌はエストロゲン作用の亢進がリスクファクターのひとつであり、乳癌同様に組織中のエストロゲン合成酵素(アロマターゼ)の発現によるパラクライン・オートクライン機構を介する癌細胞増殖促進の可能性が考えられる。我々は子宮体癌におけるアロマターゼ発現による癌の増殖・進展の機序の解明を目的とした。【方法】患者の同意を得て採取した子宮体癌組織85例について抗アロマターゼ抗体を用いて免疫染色を行い、上皮・間質・筋層に分類して発現の有無を検討した。臨床病理学的因子は進行期・筋層浸潤・組織grade・年齢等に関して統計解析を行い、Kaplan-Meier法による生存分析も行った。一部症例についてPNAプローブを用いたISH法およびlight cyclerを用いた定量的RT-PCR法にてアロマターゼmRNAの発現を定量化した。また子宮体癌由来細胞株の培養を行いアロマターゼ阻害剤を添加してその増殖抑制効果の検討を行った。【成績】臨床検体での検討では、アロマターゼは上皮52%、間質64%、筋層41%と高率に発現を認めた。上皮での発現は、筋層浸潤あり46%、なし73%、grade1 58%、grad2,3 32%とそれぞれ予後良好群に多く認められた(p=0.05,p=0.03)。筋層で発現する場合、予後不良であった(Wilcoxon p=0.05)。アロマターゼ蛋白とmRNAの発現は良く相関した。Ishikawa細胞株を用いたWST-1 assayでは、アロマターゼ阻害剤であるTZAの添加により濃度依存的に細胞増殖抑制効果を認めた。【結論】アロマターゼ発現は子宮体癌細胞の増殖に関与し、特に筋層での発現が予後と相関することが示された。筋層のアロマターゼ発現は腫瘍の筋層浸潤を促進する可能性が示唆された。研究課題/領域番号:14770842, 研究期間(年度):2002-2003出典：「子宮体癌に対するアロマターゼ遺伝子をターゲットとした遺伝子治療への戦略」研究成果報告書　課題番号14770842（KAKEN：科学研究費助成事業データベース（国立情報学研究所））（https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-14770842/)を加工して作

Performing a sperm DNA fragmentation test in addition to semen examination based on the WHO criteria can be a more accurate diagnosis of IVF outcomes

Abstract Background We analyzed the sperm DNA fragmentation index (DFI) and general semen test based on the World Health Organization (WHO) criteria and compared the two tests using semen factors. In addition, we examined whether DFI is a reliable parameter associated with in vitro fertilization (IVF) outcomes. Methods Sperm chromatin dispersion (SCD) and general semen tests were conducted in accordance with the WHO 2010 guidelines, and correlations between the two tests were investigated. The WHO criteria were set as the cutoff values for each of the following factors: semen volume, concentration, total sperm count, motility, and normal morphology, and compared with the DFI results. Results The subjects had a mean sperm DFI of 15.3% ± 12.6%, and the DFI increased with age. In contrast, motility and normal morphology decreased as the DFI increased. Patients who satisfied the WHO criteria in terms of concentration, total sperm count, and motility had a significantly lower DFI than those who did not satisfy the criteria. Therefore, evaluation with a general semen test based on the WHO criteria should be regarded as a qualitative evaluation of all factors other than semen volume and normal morphology. Conclusions High DFI (≥ 30%) caused a low blastocyst development rate following intracytoplasmic sperm injection. Male infertility due to DFI should be suspected when IVF results are poor despite normal semen findings based on the WHO criteria. The results of this study suggest that the SCD test may more accurately evaluate the correlation between IVF clinical outcomes and male infertility. Therefore, it is important to focus on DFI measurements

Managing Ectopic Pregnancies by Targeting Chorionic Villi with a Transvaginal Injection of Ethanol into the Lacunar Space

Methotrexate has been the main mode of non-surgical treatment for ectopic pregnancies. However, we have developed an easier, repeatable method that can be applied even to patients with a high beta-human chorionic gonadotropin (β-hCG) level and/or positive fetal heartbeat, by targeting chorionic villi with a transvaginal injection of absolute ethanol (AE) into the lacunar space (intervillous space). The efficacy and safety of this method were examined in 242 cases of ectopic pregnancy, including 103 with positive fetal heartbeat. Serum β-hCG level was measured at frequent intervals, and transvaginal ultrasonography was performed to observe the gestational sac and hyperechoic inner ring. Of the 242 patients, 222 (91.7%) were successfully treated. The average number of AE injection(s) required was 1.6 (range: 1–5), and the average dose was 3.2 mL. After the treatment, many of the patients tried to conceive again, and 63 of the traceable 145 patients (43.4%), who had fallopian tube pregnancy, and 7 of the traceable 12 patients (58.3%), who had cervical or cesarean scar pregnancies, successfully conceived and delivered babies with no observed side effects. Therefore, this method could be an effective treatment for ectopic pregnancy with the potential to replace conventional surgical interventions and medical treatment using methotrexate

Minimal ovarian stimulation combined with elective single embryo transfer policy: age-specific results of a large, single-centre, Japanese cohort

Abstract Background The two main complications associated with the use of assisted reproduction techniques, ovarian hyperstimulation syndrome and multiple pregnancies, could be eliminated by milder ovarian stimulation protocols and the increased use of a single embryo transfer (SET) policy. A retrospective, cohort study was performed in private infertility centre to evaluate the embryological and clinical results of a large exclusively SET program according to patient age (lower or equal 29, 30–34, 35–39, 40–44 and equal or higher 45 years). Materials A total of 7,244 infertile patients have undergone 20,244 cycles with a clomiphene-based minimal stimulation or natural cycle IVF protocol during 2008. Following oocyte retrieval, fertilization and embryo culture a total of 10,401 fresh or frozen single embryo transfer procedures were performed involving cleavage-stage embryos or blastocysts. Results Successful oocyte retrieval rate (78.0 %) showed no age-dependent decrease until 45 years. Fertilization (80.3 %) and cleavage (91.1 %) rates were not significantly different between age groups. Blastocyst formation (70.1 % to 22.8 %) and overall live birth rates (35.9 % to 2 %) showed an age-dependent decrease. Frozen-thawed blastocyst transfer cycles gave the highest chance of live birth per embryo transfer (41.3 % to 6.1 %). Conclusions High fertilization and cleavage rates were obtained regardless of age whereas blastocyst formation and live birth rates showed an age-dependent decrease. An elective single embryo transfer program based on a minimal ovarian stimulation protocol yields acceptable live birth rates per embryo transfer in infertile patients up until their mid-forties. However in very advanced age patients (equal or higher 45 years old) success rates fall below 1 %.</p