The investigation of the death-inducing potency of a recombinant Adenovector expressing Mda-7-tlyp-1 on different cancer cell lines.

Q3 Medicine Gastroenterology and Hepatology From Bed to Bench Pub Date : 2024-01-01 DOI:10.22037/ghfbb.v17i1.2779

Fatemeh Vatanparast, Rozita Ghojoghi, Maryam Kadkhodazadeh, Fatemeh Nekooei, Kazem Baesi, Mahroo Rastegari, Fatemeh Jamali, Zahra Farmani, Jamal Sarvari, Seyed Younes Hosseini

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Abstract

Aim: The potency of Adenovector expressing Mda7-tLyp1 (Ad-Mda7-tLyp1) for death induction was evaluated on the breast (MCF7), liver (HepG2), and gastric (MKN45) cancer cell lines.

Background: Mda-7 could be a possible complementary to traditional cancer therapy, and tethering to tumor-homing peptides (THPs) might improve its therapeutic efficacy.

Methods: After the preparation of recombinant Ad-Mda7-tLyp1 and Ad-Mda7, the expression of recombinant proteins was analyzed by ELISA. Adenovectors were transduced (MOI=2-5) into Hep-G2, MCF7, MKN45, and normal skin fibroblast, then tumor-killing effect was measured by cytopathic effect (CPE) monitoring, MTT viability test, BAX gene expression analysis, and Caspase3/7 assay.

Results: ELISA assay revealed a sustained level of recombinant protein secretion following Adenovector transduction. In CPE microscopy, all cancer cell lines showed a significant reduction (≥50%) in their normal phenotype after receiving Ad-Mda7-tLyp1 and Ad-Mda7. The viability was significantly lower compared to the control, indicating an anti-proliferating effect. In parallel, the viability test showed that Ad-Mda7 and Ad-Mda7-tLyp1 have a significant killing effect (≥50%) on MCF-7, Hep-G2, and MKN45 compared to normal fibroblast (P≤0.05). BAX gene expression analysis showed that both Ad-Mda7-tLyp1 and Ad-Mda7 vectors induced >2-fold increase of apoptosis (P<0.05), particularly in MCF7. Similarly, caspase3/7 activity showed a significant increase (P<0.05) following Ad-Mda7, and Ad-Mda7-tLyp1 transduction into cancer cell lines, but not in normal fibroblasts.

Conclusion: The newly constructed Ad-Mda-tlyp1 showed a suitable tumor cell killing activity and enough specificity on studied cell lines.

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研究表达 Mda-7-tlyp-1 的重组 Adenovector 对不同癌细胞株的致死效力。

目的：评估了表达Mda7-tLyp1（Ad-Mda7-tLyp1）的Adenovector在乳腺癌（MCF7）、肝癌（HepG2）和胃癌（MKN45）细胞系中诱导死亡的效力：背景：Mda-7可能是传统癌症疗法的一种补充，与肿瘤归巢肽（THPs）配伍可能会提高其疗效：方法：制备重组 Ad-Mda7-tLyp1 和 Ad-Mda7 后，用 ELISA 分析重组蛋白的表达。将腺病毒载体（MOI=2-5）转导到Hep-G2、MCF7、MKN45和正常皮肤成纤维细胞中，然后通过细胞病理效应（CPE）监测、MTT活力测试、BAX基因表达分析和Caspase3/7检测来测定肿瘤杀伤效果：结果：酶联免疫吸附试验（ELISA）显示，腺病毒载体转导后，重组蛋白分泌水平持续上升。在 CPE 显微镜下，所有癌细胞系在接受 Ad-Mda7-tLyp1 和 Ad-Mda7 后，其正常表型均显著减少（≥50%）。与对照组相比，细胞存活率明显降低，这表明存在抗增殖作用。同时，活力测试表明，与正常成纤维细胞相比，Ad-Mda7 和 Ad-Mda7-tLyp1 对 MCF-7、Hep-G2 和 MKN45 有明显的杀伤作用（≥50%）（P≤0.05）。BAX基因表达分析表明，Ad-Mda7-tLyp1和Ad-Mda7载体都能诱导细胞凋亡增加2倍以上（PConclusion：新构建的 Ad-Mda-tlyp1 对所研究的细胞株具有适当的肿瘤细胞杀伤活性和足够的特异性。

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Gastroenterology and Hepatology From Bed to Bench Medicine-Gastroenterology

CiteScore

2.30

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0.00%

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