Kazuyuki Fujita

Prostate-specific and Androgen-dependent Expression of a Novel Homeobox Gene

A new member of the mouse NK family of homeobox genes that is related to Drosophila NK-3 has been identified. Expression of this gene, termed Nkx-3.1, is largely restricted to the prostate gland in adult animals. The level of Nkx-3.1 mRNA decreases markedly in response to castration, suggesting that its expression is androgen-dependent. In situ hybridization analyses demonstrated that expression of Nkx-3.1 in the prostate is confined to epithelial cells. In newborns, Nkx-3.1 mRNA is detected in the urethral epithelium that is being induced by the surrounding mesenchyme to invaginate to form prostatic buds. Together, these observations suggest that the Nkx-3.1 protein, which likely functions as a transcription factor, plays a prominent role both in the initiation of prostate development and in the maintenance of the differentiated state of prostatic epithelial cells.

Androgen-independent expression of hoxb-13 in the mouse prostate.

Hox genes encode transcriptional regulatory proteins that are largely responsible for establishing the body plan of all metazoan organisms. A subset of Hox genes is expressed during the period of organogenesis and into adulthood. hoxb‐13 is a recently‐described member of the Hox gene family that is expressed in the spinal cord, hindgut, and urogenital sinus during embryogenesis.

Seminal plasma inhibin‐B level is a useful predictor of the success of conventional testicular sperm extraction in patients with non‐obstructive azoospermia

Aim: The value of serum inhibin‐B as a predictor of the presence of testicular spermatozoa is still controversial. The purpose of this study is to evaluate the predictive value of the seminal plasma inhibin‐B level, which might more directly reflect the secretion by Sertoli cells, and to discriminate between successful and failed testicular sperm extraction (TESE) in non‐obstructive azoospermia.

Growth inhibition of human glioma cells modulated by retrovirus gene transfection with antisense IL-8

The human glioma cell line, NP-1, expresses IL-8 mRNA and constitutively secretes IL-8 protein. Administration of recombinant IL-8 increased the proliferation of NP-1 cells in a dose-dependent manner. In the presence of anti-IL-8 antiserum, the IL-8 mediated proliferation of NP-1 cell growth was inhibited. Further, NP-1 cell growth was inhibited by transfection of retroviral constructs encoding antisense IL-8 bothin vitro andin vivo models. These results suggest that antisense IL-8 gene therapy could be beneficial in gliomas where autocrine stimulation by IL-8 is implicated.

Analysis of the complications after radical hysterectomy for stage IB, IIA and IIB uterine cervical cancer patients.

Aim:  This study was undertaken to assess whether radical hysterectomy and pelvic lymphadenectomy could be carried out within acceptable complications in uterine cervical cancer patients.

Suppression of TGF-β1 in Human Gliomas by Retroviral Gene Transfection Enhances Susceptibility to LAK Cells

Human glioma cell line, Onda 10 produces TGF-β1. TGF-β1 has a biological role for the immunosuppression of the host. We have investigated whether suppression of TGF-β1 on human glioma cell enhanced the susceptibility to lymphokine-activated killer (LAK) cells. In vitro, susceptibility to LAK cells on Onda 10 cell is augmented by retroviral gene transfection with antisense TGF-β1. Nude mice bearing Onda 10 cells transduced with antisense TGF-β1 gene has a longer life span compared to mice carrying that of sense TGF-β1 gene or vector alone. The cytotoxic activity of LAK cells induced from spleen cells of mice carrying antisense TGF-β1 gene transduced cells is higher against Onda 10 cell than that of LAK cells from mice carrying vector alone transduced cells. Also, antisense TGF-β1 gene transduced cells are much more sensitive to LAK cells compared to Onda 10. These suggest that the augmented host systemic immunity in mice is one of the mechanisms of the reduced tumorigenicity of antisense TGF-β1 gene transduced cells and that the increased systemic immunity could be ascribed to the increased immunogenicity of the tumor cells. The gene therapy for malignant glioma with antisense TGF-β1 gene is expected to be promising.

Direct Intratumoral Gene Transfer of the Herpes Simplex Virus Thymidine Kinase Gene with DNA‐liposome Complexes: Growth Inhibition of Tumors and Lack of Localization in Normal Tissues

To constitute the site‐specific expression of the herpes simplex virus thymidine‐kinase (HSV‐TK) gene in tumor cells, we have assessed the promoter function of the simian virus 40 (SV40) promoter and the 5’flanking region of c‐erbB‐2 gene using a luciferase‐expressing reporter plasmid. After the transfec‐tion of the luciferase plasmid directed by the promoter region of c‐erbB‐2 gene, a large amount of luciferase activity was observed in c‐erbB‐2‐expressing cells (Colo201, MCF‐7, and HEC1‐A), while none was detected in cells with no expression of c‐erbB‐2 protein (HRA and KF cells). On the other hand, a high level of luciferase activity was detected in all tumor cell lines tested, when the transfection was performed with SV40 promoter. The repeated transfection of the liposome‐conjugated HSV‐TK gene regulated by the SV40 promoter or by the promoter region of c‐erbB‐2 gene with cultivation in 100 μg/ml of aciclovir for 5 days in vitro resulted in growth inhibition for all four cell lines examined or for only c‐erbB‐2‐expressing cells in the presence of SV40 promoter or c‐erbB‐2 promoter, respectively. Finally, direct injection of the DNA‐liposome complex into established tumors in the presence of 50 mg/kg of aciclovir led to significant tumor volume reduction in all three tumors tested when SV40 promoter was employed. However, this anti‐tumor effect was noted only in c‐erbB‐2‐positive cells (Colo201 cells) upon intratumoral injection of HSV‐TK gene regulated by c‐erbB‐2 promoter. In the case of intratumoral gene transfer, foreign DNA was detected in only one of seven mice by polymerase chain reaction (PCR) analysis performed 7 days following injection. When PCR analysis was carried out at 14 or 21 days following injection, no DNA signal was found at all. However, DNA was detected in several normal tissues at all three times tested in the case of intravenous injection. No abnormalities were seen in histologic examinations of normal tissues or in serum biochemical parameters following DNA liposome delivery. These results suggest that the direct gene transfer of HSV‐TK gene regulated by tumor‐specific transcriptional units may be one of the most clinically promising of the selective genetic strategies against cancer.

Outcomes of fertility‐sparing surgery for women of reproductive age with FIGO stage IC epithelial ovarian cancer

To report the clinical outcomes of patients with FIGO stage IC epithelial ovarian cancer (EOC) treated by fertility‐sparing surgery (FSS).

Primary retroperitoneal mucinous cystadenocarcinoma associated with pregnancy

Primary retroperitoneal mucinous cystadenocarcinoma (PRMC) is an extremely rare tumor. Only 30 cases have been reported previously in the English literature, and little information is available concerning its treatment and prognosis. The patient was a 28-year-old woman, presenting with a right mid-abdominal tumor at 26 weeks of gestation. At 31 weeks of gestation, she underwent an exploratory laparotomy and was diagnosed with a PRMC. No disseminated tumor was observed, and an excision of only the tumor was performed. She had an uneventful vaginal delivery at 38 weeks of gestation and remains free of disease at 13 months after the operation. This report describes a case of PRMC associated with pregnancy. The optimal management of these retroperitoneal masses during pregnancy is discussed. Based on limited experience and the current literature, a PRMC with an intact capsule and no dissemination appears to have a good prognosis and can be treated by tumor excision alone in patients who wish to preserve fertility

Stage IVa squamous cell carcinoma of the vulva managed with primary chemoradiation.

We present the case of a 53-year-old woman with International Federation of Gynecology and Obstetrics stage IVa (T3N2M0) squamous cell carcinoma of the vulva. Because the urethra was surrounded by a vulvar tumor, she was managed with primary chemoradiation in an attempt to spare the morbidity associated with exenterative vulvar surgery. Treatment was given as a planned split course, consisting of two separate courses of 23.8 Gy each. During each split course of radiation, 5-fluorouracil, 1000 mg/m2 per day, was given over the first 4 days, and cisplatin 50 mg/m2 was administered as a single infusion on the first day. During the 4 days of chemotherapy infusion, the radiation was administered in two daily fractions of 1.7 Gy each, given at least 6 h apart. There was no treatment break due to adverse effect, and a pathological complete response was achieved in the primary tumor and the lymph nodes. The patient did not undergo surgical intervention, and has had no evidence of recurrence for 24 months. Chemoradiation therapy should be considered as an option in patients with locally advanced vulvar cancer to avert the need for exenterative surgery, and to preserve sexual, gastrointestinal, and urinary function.