title: Selective suppression of autocatalytic caspase-3 driven by two-step transcriptional amplified human telomerase reverse transcriptase promoter on ovarian carcinoma growth in vitro and in mice.
abstract: The objective of our study was to construct recombinant adenovirus (rAd) AdHTVP2G5-rev-casp3, which expresses autocatalytic caspase-3 driven by human telomerase reverse transcriptase promoter (hTERTp) with a two-step transcription amplification (TSTA) system and investigate its antitumor effects on ovarian cancer in vitro and in vivo. Fluorescent detection was used to detect EGFP expression in various cells. Cell viabilities were determined using the Cell Counting Kit-8 and flow cytometry. RT-PCR and immunoblotting assays were used to detect cellular apoptotic activities. Tumor growth and survival of tumor-bearing mice were studied. The hTERTp-TSTA system showed the strongest activity in hTERT-positive cancer cells when compared with hTERTp and cytomeglovirus promoter (CMVp). In contrast, it showed no activity in hTERT‑negative HUVECs. AdHTVP2G5‑rev-casp3 markedly suppressed the survival of AO cells in a dose-dependent modality with a viability rate of 17.8 ± 3.5% at an MOI of 70, which was significantly lower than that by AdHT-rev-casp3 and Ad-rev-casp3 (rAds which express rev-caspase-3 driven by hTERTp and CMVp, respectively). In contrast, AdHTVP2G5‑rev-casp3 induced little HUVEC death with a viability rate of 92.7 ± 5.2% at the same MOI. Additionally, AdHTVP2G5-rev-casp3 (MOI=70) caused significant apoptosis in AO cells with an apoptotic rate of 42%. The tumor growth suppression rate of AdHTVP2G5-rev-casp3 was 81.52%, significantly higher than that of AdHT-rev-casp3 (54.94%) or Ad-rev-casp3 (21.35%). AdHTVP2G5-rev-casp3 significantly improved the survival of tumor-bearing mice with little liver damage, with a mean survival of 258 ± 28 days. These results showed that AdHTVP2G5-rev-casp3 caused effective apoptosis with significant tumor selectivity, strongly suppressed tumor growth and improved mouse survival with little liver toxicity. It can be a potent therapeutic agent for tumor targeted treatment of ovarian cancer.
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, P.R. China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Song
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, P.R. China.
authors.forename: Xing
authors.initials: X
authors.lastname: Xin
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, P.R. China.
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Xia
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, P.R. China.
authors.forename: Xingyue
authors.initials: X
authors.lastname: Zhai
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, P.R. China.
authors.forename: Keng
authors.initials: K
authors.lastname: Shen
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000235:genetics* / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003587:Cytomegalovirus / Q000302:isolation & purification; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D009374:Neoplasms, Experimental; D009928:Organ Specificity; D010051:Ovarian Neoplasms / Q000473:pathology / Q000628:therapy*; D011401:Promoter Regions, Genetic; D016019:Survival Analysis; D019098:Telomerase / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
chemical_list: D019098:Telomerase; D053148:Caspase 3