Nakamura Hitomi, Kimura Tadashi, Ikegami Hiroyuki, Ogita Kazuhide, Koyama Shinsuke, Shimoya Koichiro, Tsujie Tomoko, Koyama Masayasu, Kaneda Yasufumi, Murata Yuji
Division of Obstetrics and Gynecology, Department of Specific Organ Regulation, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Mol Hum Reprod. 2003 Oct;9(10):603-9. doi: 10.1093/molehr/gag079.
The uterus is obviously critical in implantation, development of the fetus and parturition. Endometrial cancer derived from endometrial epithelium is one of the common malignancies in the female reproductive tract. In order to clarify the local mechanisms of reproductive physiology and establish a non-systemic therapeutic strategy for reproductive failure as well as for endometrial cancer, we applied haemagglutinating virus of Japan envelope (HVJ-E) vector to in-vivo gene transfer into the uterine cavity of IVCS mice. Injection of HVJ-E vector into mouse uterine cavity on day 1.5 post coitum (p.c.) introduced a reporter gene approximately 120-fold more efficiently than introduction using the cationic liposome method. The expression of the introduced gene continued for at least 3 days. The plasmid vector was localized in the endometrial epithelium, whereas oligo deoxynucleotides were distributed throughout the epithelium, stromal cells and myometrium. HVJ-E vector did not affect the pregnancy rate, course of pregnancy, litter size, fetal growth in utero or parturition, and did not transfect the exogenous gene to the fetus. These results indicate that gene transfer into the uterus using HVJ-E vector is highly efficient and safe during pregnancy, and results in a well controlled distribution of the exogenous DNA. We believe that this procedure should be widely applicable for investigations of reproductive physiology as well as for methods of local gene therapy in the uterus.
子宫在着床、胎儿发育和分娩过程中显然至关重要。源自子宫内膜上皮的子宫内膜癌是女性生殖道常见的恶性肿瘤之一。为了阐明生殖生理的局部机制，并为生殖功能衰竭以及子宫内膜癌建立一种非全身性的治疗策略，我们将日本血凝病毒包膜（HVJ-E）载体应用于向IVCS小鼠子宫腔内进行体内基因转移。在交配后第1.5天（p.c.）将HVJ-E载体注入小鼠子宫腔，其导入报告基因的效率比使用阳离子脂质体方法高出约120倍。导入基因的表达持续至少3天。质粒载体定位于子宫内膜上皮，而寡脱氧核苷酸则分布于整个上皮、基质细胞和肌层。HVJ-E载体不影响妊娠率、妊娠过程、产仔数、子宫内胎儿生长或分娩，也不会将外源基因转染至胎儿。这些结果表明，在妊娠期间使用HVJ-E载体向子宫内进行基因转移高效且安全，并能使外源DNA分布得到良好控制。我们认为，该方法应广泛应用于生殖生理研究以及子宫局部基因治疗方法。
