Yoshida Masanori, Yamamoto Naritaka, Nitta Takashi, Uehara Tetsuya, Terao Ryuta, Hatano Etsuro, Iimuro Yuji, Yamaoka Yoshio
Department of Gastroenterological Surgery, Kyoto University, Kyoto 606-8507, Japan.
J Surg Res. 2002 Feb;102(2):95-101. doi: 10.1006/jsre.2001.6301.
Proliferative cholangitis (PC) associated with hepatolithiasis results in stricture of the main bile ducts and is a major cause of residual and/or recurrent stones after repeated treatment for hepatolithiasis. The transcription factor E2F controls the expression of several genes involved in cell proliferation. The aim of this study was to inhibit PC using cytostatic gene therapy by transferring fusigenic anionic liposome-hemagglutinating virus of Japan (HVJ-anionic liposome) complexes containing a synthetic double-stranded oligodeoxynucleotide with high affinity for E2F (E2F decoy).
PC was induced by introducing a fine nylon thread into the bile duct in a rat model. HVJ-anionic liposomes containing the E2F decoy were administered directly into the biliary tract. HVJ-anionic liposomes containing a missense oligodeoxynucleotide (scramble decoy) were also given as a control. The count of peribiliary glands in the bile duct, 5'-bromodeoxyuridine (BrdU) labeling index, and immunohistochemical staining for proliferating cell nuclear antigen (PCNA) in the bile duct were compared among untransfected, scramble decoy-transfected, and E2F decoy-transfected rats.
E2F decoy-transfected bile ducts showed inhibition of the papillary proliferation of the biliary epithelium and peribiliary gland hyperplasia. BrdU incorporation and PCNA expression in the bile ducts were inhibited in E2F decoy-transfected rats.
Our cytostatic gene therapy approach using direct E2F decoy transfer into the biliary tract suppressed PC in a rat model and may offer an effective therapeutic option for reducing recurrence following treatment for hepatolithiasis.
与肝内胆管结石相关的增殖性胆管炎（PC）会导致肝内主要胆管狭窄，是肝内胆管结石反复治疗后结石残留和/或复发的主要原因。转录因子E2F控制着多个参与细胞增殖的基因的表达。本研究的目的是通过转染含有对E2F具有高亲和力的合成双链寡脱氧核苷酸（E2F诱饵）的融合阴离子脂质体 - 日本血凝病毒（HVJ - 阴离子脂质体）复合物，利用细胞生长抑制基因疗法抑制PC。
在大鼠模型中，通过将细尼龙线引入胆管来诱导PC。将含有E2F诱饵的HVJ - 阴离子脂质体直接注入胆道。还给予含有错义寡脱氧核苷酸（随机诱饵）的HVJ - 阴离子脂质体作为对照。比较未转染、随机诱饵转染和E2F诱饵转染大鼠胆管周围腺体的数量、5'-溴脱氧尿苷（BrdU）标记指数以及胆管中增殖细胞核抗原（PCNA）的免疫组化染色情况。
E2F诱饵转染的胆管显示出胆管上皮乳头样增殖和胆管周围腺体增生受到抑制。E2F诱饵转染的大鼠胆管中BrdU掺入和PCNA表达受到抑制。
我们将E2F诱饵直接导入胆道的细胞生长抑制基因治疗方法在大鼠模型中抑制了PC，可能为减少肝内胆管结石治疗后的复发提供一种有效的治疗选择。
