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Reduced RAR-β gene expression in Benzo(a)Pyrene induced lung cancer mice is upregulated by DOTAP lipo-ATRA treatment
S Viswanathan1, V M Berlin Grace1
1Department of Biotechnology, Karunya Institute of Technology and Sciences, Karunya Nagar, Coimbatore 641 114, Tamil Nadu, India.
Abstract:
Molecular targeted therapy for specific genes is an emerging research. Retinoic Acid Receptor (RAR-β) is a key tumor suppressor which is found to be lost drastically during much cancer progression. We hence, analyzed the expression level of RAR-β gene during B(a)P induced lung cancer development in mice and studied the lung cancer targeted action of All Trans Retinoic Acid (ATRA) in DOTAP liposomal formulation. The effect of its treatment on lung cancer was determined by histopathological analysis. RAR-β gene expression was assessed by RT-PCR and qPCR. A distinct band for RAR-β gene (density - 0.5123 for lung and 0.5160 for liver) was observed in normal mice, whereas no visible band was observed in cancer induced group, indicating loss of RAR-β gene expression. Both ATRA and lipo-ATRA treated groups showed detectable RAR-β expression with relatively lesser density than the normal group. The expression was more intense in lipo-ATRA treatment (density-0.2973) compared with free ATRA treatment (density-0.1549) in lung tissues. The qPCR results also have highlighted a highly significant (p ≤ 0.01) variation RQ values between lipo-ATRA group (15.46 ± 1.54) and free ATRA group (7.58 ± 1.30) in lung tissue sample on 30th day. The mean RQ value for normal lung on 30th day was 20.86 ± 2.58 against the cancer control. The 120th day mice also showed the similar RAR-β expression pattern with further declined expression levels as there was no treatment given after 30 days. Interestingly, the lipo-ATRA treatment could show a highly significant (p ≤ 0.001) expression (12.00 ± 2.31) when compared with free ATRA treatment (3.31 ± 0.58) which implies that the lipo-ATRA formulation could result in sustained delivery of ATRA in target site. Histopathology of lung and liver on 120th day also revealed an effective therapeutic indication in lipo-ATRA treatment compared to free ATRA treatment due to lipo-ATRA's stealth property and it efficiently inhibited the metastasis to liver. These results revealed that the lipo-ATRA treatment has efficiently delivered ATRA into target site than free ATRA and in-turn it might have induced the expression of RAR-β gene or prevented loss of RAR-β gene in cancer animals.
Insights
This study shows that liposomal All Trans Retinoic Acid (lipo-ATRA) effectively delivers the drug to lung cancer cells, restoring Retinoic Acid Receptor-beta (RAR-β) expression and inhibiting tumor spread. Lipo-ATRA demonstrates superior therapeutic potential compared to free ATRA.
Area of Science:
- Oncology
- Molecular Biology
- Drug Delivery Systems
Background:
- Retinoic Acid Receptor-beta (RAR-β) is a crucial tumor suppressor gene often lost in cancer.
- Benzo[a]pyrene (B[a]P) is a known carcinogen that induces lung cancer in animal models.
- All Trans Retinoic Acid (ATRA) is a therapeutic agent, but its targeted delivery remains a challenge.
Purpose of the Study:
- To investigate the expression levels of RAR-β during B[a]P-induced lung cancer in mice.
- To evaluate the efficacy of ATRA delivered via DOTAP liposomal formulation (lipo-ATRA) for lung cancer treatment.
- To compare the therapeutic effects of lipo-ATRA with free ATRA.
Main Methods:
- Lung cancer was induced in mice using B[a]P.
- RAR-β gene expression was analyzed using Reverse Transcription Polymerase Chain Reaction (RT-PCR) and quantitative PCR (qPCR).
- Histopathological analysis of lung and liver tissues was performed to assess treatment effects and metastasis.
- ATRA was administered both as a free drug and in a DOTAP liposomal formulation.
Main Results:
- RAR-β gene expression was significantly reduced in B[a]P-induced lung cancer compared to normal mice.
- Both free ATRA and lipo-ATRA treatments restored RAR-β expression, with lipo-ATRA showing more intense expression.
- qPCR analysis revealed significantly higher RAR-β expression in the lipo-ATRA group compared to the free ATRA group.
- Histopathology confirmed better therapeutic outcomes and reduced liver metastasis with lipo-ATRA treatment, indicating sustained drug delivery and efficacy.
Conclusions:
- Lipo-ATRA formulation provides sustained delivery of ATRA to the target site, enhancing its therapeutic effect in lung cancer.
- The study highlights the potential of lipo-ATRA as an effective molecular targeted therapy for lung cancer by restoring RAR-β expression.
- DOTAP liposomal encapsulation improves ATRA's efficacy and reduces off-target effects, such as liver metastasis.
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