Ameliorating effect of lipo-ATRA treatment on the expression of TIG3 and its suppressing effect on PPARγ gene expression in lung cancer animal model

  • Ragavi Ravichandran
  • S. Viswanathan
  • V. M. Berlin GraceEmail author
  • Lucia Bonati
  • Jini Narayanan


This study aimed to find out the molecular therapeutic effect of lipo-ATRA on tumour suppressor TIG3 and cell proliferative biomarker PPARγ in B (a) P-induced lung cancer model. In RT-PCR study, ATRA- and lipo-ATRA-treated mice samples showed relatively higher TIG3 expression and decreased PPARγ expression (Band density) than cancer control. Among treatments, lipo-ATRA showed vital effect than free ATRA by enhancing TIG3 and decreasing PPARγ. The qPCR results also showed significant (p ≤ 0.05) difference in both TIG3 and PPAR (RQ values of TIG3, lipo-ATRA 23.85 ± 1.29; free ATRA 10.43 ± 1.81 and for PPARγ, lipo-ATRA 4.707 ± 1.21; free ATRA 15.78 ± 2.34). From this, we conclude that liposomal ATRA formulation is most preferable for prolonged delivery of ATRA at targeted site to favour molecular action. It implies that the therapeutic effect of lipo-ATRA in lung cancer was exhibited by ameliorating the TIG3 expression and by suppressing the expression of PPARγ.


Lung cancer Lipo-ATRA Benzo (a) pyrene TIG3 PPARγ 



Tazarotene-induced gene 3


Peroxisome proliferator-activated receptor gamma


Retinoic acid receptor


Retinoid X receptor


All trans retinoic acid


Retinoic acid-responsive elements

B (a) P

Benzo (a) Pyrene


1, 2-Dioleoyl-3-trimethylammonium-propane


Relative quantity



We are very grateful to thank the Department of Science and Technology-Science and Engineering Research board (DST-SERB), Govt. of India [SB/YS/LS-252/2013 (May 15, 2014)], Department of Biotechnology (DBT), Govt of India [BT/PR14632/NNT/28/824/2015] and Karunya short-term Research grant 2018–2019 [KITS/KSG/56/2018] for the financial support given to complete this work successfully. We extend our thanks to Ms. Perinba Danisha J, SRF, KITS, and Mr. P. Jeyakumar for their valuable guidance and technical assistance during this research work. We acknowledge the Karunya Institute of Technology and Sciences, Coimbatore, for providing instruments and laboratory facilities. We also thank the timely help rendered by the authorities of Sugarcane Breeding Institute, Coimbatore, to carry out the real-time qPCR for the gene expression study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    World Health Statistics (2018) Monitoring health for sustainable development goals. World Health Organization 2018Google Scholar
  2. 2.
    Toh CK (2009) The changing epidemiology of lung cancer. Cancer epidemiology. Humana Press, New York, pp 397–411. CrossRefGoogle Scholar
  3. 3.
    Pleasance ED, Stephens PJ, O’meara S S, McBride DJ, Meynert A, Jones D et al (2010) A small-cell lung cancer genome with complex signatures of tobacco exposure. Nature 463(7278):184. CrossRefGoogle Scholar
  4. 4.
    Takahashi H, Ogata H, Nishigaki R, Broide DH, Karin M (2010) Tobacco smoke promotes lung tumorigenesis by triggering IKKβ-and JNK1-dependent inflammation. Cancer Cell 17(1):89–97. CrossRefGoogle Scholar
  5. 5.
    Hecht SS (2012) Lung carcinogenesis by tobacco smoke. Int J Cancer 131(12):2724–2732. CrossRefGoogle Scholar
  6. 6.
    Kim JH, Yamaguchi K, Lee SH, Tithof PK, Sayler GS, Yoon JH, Baek SJ (2005) Evaluation of polycyclic aromatic hydrocarbons in the activation of early growth response-1 and peroxisome proliferator activated receptors. ToxicologicalSciences 85(1):585–593. Google Scholar
  7. 7.
    Shafey O, Eriksen M, Ross H, Mackay J (2009) The tobacco atlas. Atlanta 3:38–39Google Scholar
  8. 8.
    Kometani T, Yoshino I, Miura N, Okazaki H, Ohba T, Takenaka T, Maehara Y (2009) Benzo [a] pyrene promotes proliferation of human lung cancer cells by accelerating the epidermal growth factor receptor signaling pathway. Cancer Lett 278(1):27–33. CrossRefGoogle Scholar
  9. 9.
    Klaunig JE, Wang Z, Pu X, Zhou S (2011) Oxidative stress and oxidative damage in chemical carcinogenesis. Toxicol Appl Pharmacol 254(2):86–99. CrossRefGoogle Scholar
  10. 10.
    Pfeifer GP, Denissenko MF, Olivier M, Tretyakova N, Hecht SS, Hainaut P (2002) Tobacco smoke carcinogens, DNA damage and p53 mutations in smoking-associated cancers. Oncogene 21(48):7435. CrossRefGoogle Scholar
  11. 11.
    Kasala ER, Bodduluru LN, Barua CC, Sriram CS, Gogoi R (2015) Benzo (a) pyrene induced lung cancer: role of dietary phytochemicals in chemoprevention. Pharmacol Rep 67(5):996–1009. CrossRefGoogle Scholar
  12. 12.
    Miller KP, Ramos KS (2001) Impact of cellular metabolism on the biological effects of benzo [a] pyrene and related hydrocarbons. Drug Metab Rev 33(1):1–35. CrossRefGoogle Scholar
  13. 13.
    Kang L, Gao Z, Huang W, Jin M, Wang Q (2015) Nanocarrier-mediated co-delivery of chemotherapeutic drugs and gene agents for cancer treatment. Acta Pharm Sin B 5(3):169–175. CrossRefGoogle Scholar
  14. 14.
    Barr Kumarakulasinghe N, Zanwijk NV, Soo RA (2015) Molecular targeted therapy in the treatment of advanced stage non-small cell lung cancer (NSCLC). Respirology 20(3):370–378. CrossRefGoogle Scholar
  15. 15.
    Regazzi MB, Iacona I, Gervasutti C, Lazzarino M, Toma S (1997) Clinical pharmacokinetics of tretinoin. Clin Pharmacokinet 32(5):382–402. CrossRefGoogle Scholar
  16. 16.
    Ozpolat B, Lopez-Berestein G, Adamson P, Fu CJ, Williams AH (2003) Pharmacokinetics of intravenously administered liposomal all-trans-retinoic acid (ATRA) and orally administered ATRA in healthy volunteers. J Pharm Pharm Sci 6(2):292–301Google Scholar
  17. 17.
    McGowan SE, Harvey CS, Jackson SK (1995) Retinoids, retinoic acid receptors, and cytoplasmic retinoid binding proteins in perinatal rat lung fibroblasts. Am J Phys 269(4):L463–L472. Google Scholar
  18. 18.
    Choi EJ, Whang YM, Kim SJ, Kim HJ, Kim YH (2007) Combinational treatment with retinoic acid derivatives in non-small cell lung carcinoma in vitro. J Korean Med Sci 22(Suppl):S52–S60. CrossRefGoogle Scholar
  19. 19.
    Siddikuzzaman GC, Berlin Grace VM (2011) All trans retinoic acid and cancer. Immunopharmacol Immunotoxicol 33(2):241–249CrossRefGoogle Scholar
  20. 20.
    Higuchi E, Chandraratna RA, Hong WK, Lotan R (2003) Induction of TIG3, a putative class II tumor suppressor gene, by retinoic acid in head and neck and lung carcinoma cells and its association with suppression of the transformed phenotype. Oncogene 22(30):4627. CrossRefGoogle Scholar
  21. 21.
    DiSepio D, Ghosn C, Eckert RL, Deucher A, Robinson N, Duvic M, Nagpal S (1998) Identification and characterization of a retinoid-induced class II tumor suppressor/growth regulatory gene. Proc Natl Acad Sci USA 95(25):14811–14815CrossRefGoogle Scholar
  22. 22.
    Klaunig JE, Babich MA, Baetcke KP, Cook JC, Corton JC, David RM, Roberts RA (2003) PPARα agonist-induced rodent tumors: modes of action and human relevance. Crit Rev Toxicol 33(6):655–780. CrossRefGoogle Scholar
  23. 23.
    Michalik L, Desvergne B, Wahli W (2004) Peroxisome-proliferator-activated receptors and cancers: complex stories. Nat Rev Cancer 4(1):61. CrossRefGoogle Scholar
  24. 24.
    Genini D, Garcia-Escudero R, Carbone GM, Catapano CV (2012) Transcriptional and non-transcriptional functions of PPARβ/δ in Non-small cell lung cancer. PLoS ONE 7(9):e46009. CrossRefGoogle Scholar
  25. 25.
    Mader S, Chen JY, Chen Z, White J, Chambon P, Gronemeyer H (1993) The patterns of binding of RAR, RXR and TR homo-and heterodimers to direct repeats are dictated by the binding specificites of the DNA binding domains. EMBO J 12(13):5029–5041. CrossRefGoogle Scholar
  26. 26.
    Inoue K, Kawahito Y, Tsubouchi Y, Yamada R, Kohno M, Hosokawa Y, Sano H (2001) Expression of peroxisome proliferator-activated receptor (PPAR)-gamma in human lung cancer. Anticancer Res 21(4A):2471–2476 (PMID:11724309) Google Scholar
  27. 27.
    Kersten S, Desvergne B, Wahli W (2000) Roles of PPARs in health and disease. Nature 405(6785):421. CrossRefGoogle Scholar
  28. 28.
    Georgiadi A, Kersten S (2012) Mechanisms of gene regulation by fatty acids. Adv Nutr 3(2):127–134. CrossRefGoogle Scholar
  29. 29.
    Grace VB, Viswanathan S (2017) Pharmacokinetics and therapeutic efficiency of a novel cationic liposome nano-formulated all trans retinoic acid in lung cancer mice model. J Drug Deliv Sci Technol 39:223–236. CrossRefGoogle Scholar
  30. 30.
    Viswanathan S, Grace VB (2018) Reduced RAR-β gene expression in Benzo (a) Pyrene induced lung cancer mice is upregulated by DOTAP lipo-ATRA treatment. Gene 668:18–26. CrossRefGoogle Scholar
  31. 31.
    Larsen JE, Minna JD (2011) Molecular biology of lung cancer: clinical implications. Clin Chest Med 32(4):703–740. CrossRefGoogle Scholar
  32. 32.
    Chang WA, Hung JY, Tsai YM, Hsu YL, Chiang HH, Chou SH, Kuo PL (2016) Laricitrin suppresses increased benzo (a) pyrene-induced lung tumor-associated monocyte-derived dendritic cell cancer progression. Oncol Lett 11(3):1783–1790. CrossRefGoogle Scholar
  33. 33.
    Chen J, Li Q (2016) Implication of retinoic acid receptor selective signaling in myogenic differentiation. Sci Rep 6:18856. CrossRefGoogle Scholar
  34. 34.
    Ramya D, Siddikuzzaman, Grace VB (2012) Effect of all-trans retinoic acid (ATRA) on syndecan-1 expression and its chemoprotective effect in benzo (α) pyrene-induced lung cancer mice model. Immunopharmacol Immunotoxicol 34(6):1020–1027. CrossRefGoogle Scholar
  35. 35.
    Huang SL, Shyu RY, Yeh MY, Jiang SY (2000) Cloning and characterization of a novel retinoid-inducible gene 1 (RIG1) deriving from human gastric cancer cells. Mol Cell Endocrinol 159(1–2):15–24. CrossRefGoogle Scholar
  36. 36.
    Mangelsdorf DJ (1994) Vitamin A receptors. Nutr Rev 52(2):S32–S44CrossRefGoogle Scholar
  37. 37.
    Han S, Roman J (2007) Peroxisome proliferator-activated receptor γ: a novel target for cancer therapeutics? Anticancer Drugs 18(3):237–244. CrossRefGoogle Scholar
  38. 38.
    Li MY, Yuan H, Ma LT, Kong AW, Hsin MK, Yip JH, Chen GG (2010) Roles of peroxisome proliferator-activated receptor–α and–γ in the Development of non-small cell lung cancer. Am J Respir Cell Mol Biol 43(6):674–683. CrossRefGoogle Scholar
  39. 39.
    Berry DC, Noy N (2009) All-trans-retinoic acid represses obesity and insulin resistance by activating both peroxisome proliferation-activated receptor β/δ and retinoic acid receptor. Mol Cell Biol 29(12):3286–3296. CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of BiotechnologyKarunya Institute of Technology and SciencesCoimbatoreIndia
  2. 2.BiotechnologyETH ZurichBaselSwitzerland
  3. 3.Department of BiotechnologySugarcane Breeding InstituteCoimbatoreIndia

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