Brüning A, Köhler T, Quist S, Wang-Gohrke S, Moebus V J, Kreienberg R, Runnebaum I B
Department of Obstetrics and Gynecology, University of Freiburg, Germany.
Hum Gene Ther. 2001 Mar 1;12(4):391-9. doi: 10.1089/10430340150504019.
Recombinant adenoviruses expressing a therapeutic gene are currently used in clinical studies for treatment of advanced ovarian cancer. We therefore tested whether the expression level of primary (CAR) and secondary adenovirus receptors (integrins) was predictive of the efficacy of adenoviral gene transfer in ovarian cancer cells. Adenoviral transduction efficiency (ATE) was determined with an E1-deleted adenovirus type 5 expressing beta-galactosidase under a CMV promoter (AdGal). ATE was studied in relationship to the expression level of both CAR (coxsackie and adenovirus receptor) and integrins. A representative sample of 25 permanent human cell lines established from advanced ovarian cancer in our laboratory and the OV-2774 cell line were tested. Overall, ATE increased with increasing titers of AdGal. At a given titer of 50 infectious units per cell, transduction efficiency varied from 6 to 94% among the individual cell lines. All cell lines expressed CAR and integrin alpha(v)beta(5), but no relation between ATE and expression level of CAR or alpha(v)beta(5) integrin was observed. In contrast, cell lines with poor ATE, despite expressing high levels of CAR, lacked expression of integrins alpha(v)beta(3) and alpha(5)beta(1). Reconstitution of alpha(v)beta(3) integrin by reexpressing the beta(3) subunit significantly enhanced ATE of ovarian cancer cells. In ovarian cancer, neither integrins nor CAR alone appear to be potentially useful predictive markers for ATE by serotype 5 adenovirus in clinical gene therapy. A minimum level of CAR necessary for binding of adenoviruses was observed in all tested ovarian cancer cell lines. Loss of alpha(v)beta(3) integrin is frequently associated with advanced stages of ovarian cancer and can significantly reduce ATE.
表达治疗性基因的重组腺病毒目前正用于晚期卵巢癌治疗的临床研究。因此，我们测试了主要（柯萨奇病毒和腺病毒受体，CAR）和次要腺病毒受体（整合素）的表达水平是否可预测腺病毒基因转移在卵巢癌细胞中的疗效。使用在巨细胞病毒（CMV）启动子下表达β-半乳糖苷酶的E1缺失5型腺病毒（AdGal）测定腺病毒转导效率（ATE）。研究了ATE与CAR（柯萨奇病毒和腺病毒受体）和整合素表达水平的关系。测试了从我们实验室的晚期卵巢癌建立的25种永久性人类细胞系的代表性样本以及OV - 2774细胞系。总体而言，ATE随AdGal滴度增加而增加。在每细胞50个感染单位的给定滴度下，各细胞系间的转导效率从6%到94%不等。所有细胞系均表达CAR和整合素α(v)β(5)，但未观察到ATE与CAR或α(v)β(5)整合素表达水平之间的关系。相反，尽管ATE较差的细胞系表达高水平的CAR，但缺乏整合素α(v)β(3)和α(5)β(1)的表达。通过重新表达β(3)亚基来重建α(v)β(3)整合素可显著提高卵巢癌细胞的ATE。在卵巢癌中，单独的整合素或CAR似乎都不是5型腺病毒在临床基因治疗中用于预测ATE的潜在有用标志物。在所有测试的卵巢癌细胞系中均观察到腺病毒结合所需的最低CAR水平。α(v)β(3)整合素的缺失常与卵巢癌的晚期相关，并且可显著降低ATE。
