Maintaining viability and characteristics of cholangiocarcinoma tissue by vitrification-based cryopreservation

Tumor tissue has great clinical and scientific value which relies highly on the proper preservation of primary materials. Conventional tumor tissue cryopreservation using slow-freezing method has yielded limited success, leading to significant cell loss and morphological damage. Here we report a standardized vitrification-based cryopreservation method, by which we have successfully vitrified and warmed 35 intrahepatic cholangiocarcinoma (ICC) tissues with up to 80% viability of the fresh tumor tissues. Cryopreserved ICC tissue could generate patient-derived xenografts (PDXs) with take rates of 68.2% compared to 72.7% using fresh tumor tissues. Histological and genetic analyses showed that no significant alterations in morphology and gene expression were introduced by this cryopreservation method. Our procedure may facilitate collection, long-time storage and propagation of cholangiocarcinoma or other tumor specimens for (pre)clinical studies of novel therapies or for basic research.     

α7烟碱型胆碱能受体调节胆管癌细胞化疗敏感性的相关探讨

目的：观察尼古丁对胆管癌细胞QBC939化疗敏感性影响,并初步探讨其作用靶点。方法：应用MTT法检测α7烟碱型胆碱能受体激动剂尼古丁及其阻断剂仪银环蛇毒素（α-BTX）干预后的人胆管癌细胞QBC939经5-FU处理后存活增殖能力变化,应用单克隆平板试验观察尼古丁及α银环蛇毒素对5-FU处理后细胞克隆形成率变化。结果：经5-fu处理后,尼古丁刺激组（终浓度分别为10-3g／L、10-4g／L、10-5g／L）细胞存活率分别为128％、124％、118％,细胞存活率较阴性对照组明显升高,并呈一定的浓度依赖性,而α银环蛇毒素刺激组（2ug／mL）、尼古丁α银环蛇毒素联合组细胞存活率分别为92％、94％、93％、92％。尼古丁刺激组（6．2±0．40）的克隆形成能力明显高于α银环蛇毒素刺激组（3．2±0．20）、联合组（3．2±0．20）及对照组（3．4±0．33）;结论：尼古丁可明显降低胆管癌细胞对化疗药物的敏感性,其可能是通过α7烟碱型胆碱能受体发挥化疗抵抗效应。     

Overexpression of PDGFA and its receptor during carcinogenesis of Opisthorchis viverrini-associated cholangiocarcinoma

Cholangiocarcinoma (CCA) is a crucial health problem in northeastern part of Thailand, which is caused by a combination of Opisthorchis viverrini infection and nitrosamine. A better understanding of its molecular mechanism is an important step to discover and develop the new diagnostics and therapies for CCA. To reveal the involvement of potential genes in the development of CCA, the present study investigated the expression kinetics of platelet-derived growth factor alpha (Pdgfa) and its receptor (Pdgfra) during the tumorigenesis of CCA induced by O. viverrini infection with quantitative RT-PCR, and confirmed the expression with immunohistological staining. The results showed that in the hamster model of opisthorchiasis-associated CCA, the expression of Pdgfa was increased after infection plus N-nitrosodimethylamine (NDMA) administration, reached its peak at 2 months post infection, and remained at the high level until 6 months. Similarly, the expression of Pdgfra was increased time-dependently. The positive immunostaining for PDGFA proteins was observed in the cytoplasm of epithelial tumor cells of hamster CCA. Moreover, the analysis of the expression of these genes in 10 cases of human opisthorchiasis-associated CCA showed that Pdgfa was overexpressed in 80%, and Pdgfra was overexpressed in 40% cases (> 3.0 folds, compared with the expressions of adjacent normal tissues). This result suggests that PDGFA is likely involved in the tumorigenesis of opisthorchiasis-associated CCA, and may be a promising candidate biomarker for diagnosis and treatment strategies of CCA.     

肝吸虫与胆管癌

肝吸虫病是一种重要的人兽共患寄生虫病,也是我国重点防治主要食源性寄生虫病之一。肝吸虫感染被国际癌症研究中心（IARC）确定为Ⅰ类致癌因素,其所致胆管癌是肝吸虫感染的严重后果之一。本文就目前国内外肝吸虫感染的流行病学及其诱发胆管癌的发病机制研究进展和成果做一综述,为今后学者进一步深入研究肝吸虫病提供参考资料。