Antiproliferative Effects of Recombinant Apoptin on Lung and Breast Cancer Cell Lines
Masoud Ezami Razliqi1, Gholamreaza Olad2, Rouhollah Dorostkar1
1Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Background:
Selective therapy has always been the main challenge in cancer treatments. Various non-replicative oncolytic viral systems have revealed the safety and efficacy of using viruses and these products. The aim of this paper is to examine the impact of recombinant apoptin on the proliferation of lung cancer and breast cancer cell lines.
Methods:
The present study consisted of two steps of expression of recombinant apoptin and its anti-proliferative effects on normal and cancer cells. In the first step, following bioinformatics and optimizing apoptin gene sequencing and synthesis, it was expressed using vector PET28a and E. coli BL21 (DE3). The expressed recombinant apoptin was confirmed by analytical SDSPAGE and then purified using Ni affinity chromatography. In the second step, the antiproliferative effects of recombinant apoptin on lung cancer, breast cancer and primary cell lines were determined using MTT assay.
Results:
According to the results of SDS-PAGE gel assay, recombinant apoptin was visible in the 14 kDa band. Also, the MTT assay results indicated that the antiproliferative effects of recombinant apoptin in cancer cell lines was different compared with the primary cell line, and followed a dose-dependent manner in both cell lines. The highest cytotoxicity (lowest cell viability) groups were 0.2 mg/mL in lung cancer (0.32 ± 0.015) (P<0.001), and in breast cancer (0.33 ± 0.031) (P<0.001) and 0.032 mg/mL in primary cells (0.17 ± 0.004) (P<0.01), as compared to the control groups.
Conclusion:
Our results confirmed that recombinant apoptin can induce antiproliferative effects in lung cancer and breast cancer cell lines, but not in normal monkey kidney cell line Vero; thus, it can be introduced as a promising novel specific antitumor agent after further evaluation in clinical trials.
Insights
Recombinant apoptin demonstrated significant antiproliferative effects on lung and breast cancer cell lines, showing promise as a targeted antitumor agent. This protein selectively inhibits cancer cell growth without harming normal cells, warranting further clinical investigation.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Selective cancer therapy remains a significant challenge in oncology.
- Oncolytic viral systems offer a safe and effective therapeutic approach.
- Recombinant apoptin is being investigated for its potential in cancer treatment.
Purpose of the Study:
- To evaluate the impact of recombinant apoptin on the proliferation of lung and breast cancer cell lines.
- To assess the specificity of recombinant apoptin's anti-proliferative effects on normal versus cancer cells.
Main Methods:
- Recombinant apoptin was expressed in E. coli and purified using Ni affinity chromatography.
- The anti-proliferative effects were determined using MTT assay on lung cancer, breast cancer, and primary cell lines.
- SDS-PAGE was used to confirm the expression and purification of recombinant apoptin.
Main Results:
- Recombinant apoptin was successfully expressed and purified, appearing as a 14 kDa band on SDS-PAGE.
- A dose-dependent anti-proliferative effect was observed in both lung and breast cancer cell lines.
- Recombinant apoptin exhibited higher cytotoxicity in cancer cell lines compared to the normal primary cell line.
Conclusions:
- Recombinant apoptin effectively induces anti-proliferative effects in lung and breast cancer cell lines.
- The agent demonstrated specificity, with no significant impact on the normal monkey kidney cell line (Vero).
- Recombinant apoptin shows potential as a novel, specific antitumor agent for further clinical evaluation.
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