Aloin promotes oral squamous cell carcinoma cell apoptosis and autophagy through Akt/mTOR pathway
Main Article Content
Keywords
oral squamous cell carcinoma, Aloin, proliferation, migration and invasion, apoptosis and autophagy, Akt/mTOR signal transduction pathway
Abstract
Aloin, a natural product, has demonstrated anticancer activity in a variety of cancers, but its role in oral squamous cell carcinoma (OSCC) has not been reported. In this study, we aimed to investigate the effects of Aloin on the biological functions of OSCC cells. Additionally, we evaluated the classical Akt/mTOR signal transduction pathway. CAL-27 OSCC cells were treated with Aloin for in vitro experiments. 5-bromo-2’-deoxyuridine (BrdU) and colony formation assays were applied to evaluate cell proliferation potential. Wound scratch and Transwell assays detected cell migration and invasion, respectively. Flow cytometry detected apoptosis. Autophagy was assessed by p62, Beclin-1, light chain 3 (LC3)-I and LC3-II protein expressions through Western blot test. Akt/mTOR signal pathway-related protein levels were also detected by Western blot test. Here, compared with the control group, Aloin treatment memorably reduced the number of BrdU-positive cells and colonies, and undermined the ability of cell migration and invasion. Moreover, Aloin promoted apoptosis and autophagy. Western blot test proved that the protein expression of p62 was down-regulated, while Beclin-1 and LC3-I/LC3-II protein expressions were up-regulated. Further investigations revealed that Aloin inhibited p-Akt and p-mTOR, suggesting that Aloin could block the activation of AKT/mTOR pathway. Our findings demonstrated that Aloin could inhibit the activation of the AKT/mTOR pathway to induce autophagic cell death, which could provide a novel vision for OSCC treatment.
References
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Lee, I.C. and Bae, J.S., 2021. Inhibitory effects of aloin on TGFBIp-mediated septic responses. Journal of Asian Natural Products Research 23: 189–203. 10.1080/10286020.2019.1711066
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Lei, J., Shen, Y., Xv, G., Di, Z., Li, Y. and Li, G., 2020. Aloin suppresses lipopolysaccharide-induced acute lung injury by inhibiting NLRP3/NF-kappaB via activation of SIRT1 in mice. Immunopharmacology and Immunotoxicology 42: 306–313. 10.1080/08923973.2020.1765373
Li, P., Ren, K., yin Liang, Y., kai Liu J., wen Liang, Z., et al. 2020. Aloin promotes cell apoptosis by targeting HMGB1-TLR4-ERK axis in human melanoma cells. EXCLI Journal 19: 641.
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Lin, C.W., Chin, H.K., Lee, S.L., Chiu, C.F., Chung, J.G., Lin, Z.Y., et al. 2019. Ursolic acid induces apoptosis and autophagy in oral cancer cells. Environmental Toxicology 34: 983–991. 10.1002/tox.22769
Maiuri, M.C., Zalckvar, E., Kimchi, A. and Kroemer, G., 2007. Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nature Reviews in Molecular Cell Biology 8: 741–752. 10.1038/nrm2239
McMullen, P. and McMullen, A., 1989. Ambulatory centers can be a source of liability, too. Nursing Connections 2(3): 42–44.
Quadri, M.F.A., Tadakamadla, S.K. and John, T., 2019. Smokeless tobacco and oral cancer in the Middle East and North Africa: a systematic review and meta-analysis. Tobacco Induced Diseases 17: 56. 10.18332/tid/110259
Sanchez, M., Gonzalez-Burgos, E., Iglesias, I. and Gomez-Serranillos, M.P., 2020. Pharmacological update properties of aloe vera and its major active constituents. Molecules 25: 1324. 10.3390/molecules25061324
Sharma, A., Kim, J.W. and Paeng, J.Y., 2018. Clinical analysis of neck node metastasis in oral cavity cancer. Journal of the Korean Association of Oral and Maxillofacial Surgeons 44: 282–288. 10.5125/jkaoms.2018.44.6.282
Sun, R., Zhai, R., Ma, C. and Miao, W., 2020. Combination of aloin and metformin enhances the antitumor effect by inhibiting the growth and invasion and inducing apoptosis and autophagy in hepatocellular carcinoma through PI3K/AKT/mTOR pathway. Cancer Medicine 9: 1141–1151. 10.1002/cam4.2723
Tandon, P., Dadhich, A., Saluja, H., Bawane, S. and Sachdeva, S., 2017. The prevalence of squamous cell carcinoma in different sites of oral cavity at our rural health care centre in Loni, Maharashtra—a retrospective 10-year study. Contemporary Oncology (Poznan, Poland) 21: 178–183. 10.5114/wo.2017.68628
Tao, H., Tang, T., Wang, S., Wang, Z., Ma, Y., Cai, T., et al. 2019. The molecular mechanisms of aloin induce gastric cancer cells apoptosis by targeting high mobility group box 1. Drug Design, Development and Therapy 13: 1221. 10.2147/DDDT.S201818
Torre, L.A., Bray, F., Siegel, R.L., Ferlay, J., Lortet-Tieulent, J. and Jemal, A., 2015. Global cancer statistics, 2012. Cancer Journal for Clinicians 65: 87–108. 10.3322/caac.21262
Wang, Z., Tang, T., Wang, S., Cai, T., Tao, H., Zhang, Q., et al. 2020b. Aloin inhibits the proliferation and migration of gastric cancer cells by regulating NOX2-ROS-mediated pro-survival signal pathways. Drug Design, Development and Therapy 14: 145–155. 10.2147/DDDT.S219247
Wang, J., Wang, S., Wang, Y., Wang, L., Xia, Q., Tian, et al. 2020a. Chemopreventive effect of modified zengshengping on oral cancer in a hamster model and assessment of its effect on liver. Journal of Ethnopharmacology 255: 112774. 10.1016/j.jep.2020.112774
Wei, M., Wu, Y., Liu, H. and Xie, C., 2020. Genipin induces autophagy and suppresses cell growth of oral squamous cell carcinoma via PI3K/AKT/MTOR pathway. Drug Design, Development and Therapy 14: 395–405. 10.2147/DDDT.S222694
White, E., 2012. Deconvoluting the context-dependent role for autophagy in cancer. Nature Reviews Cancer 12: 401–410. 10.1038/nrc3262
Xia, P. and Xu, X.Y., 2015. PI3K/Akt/mTOR signaling pathway in cancer stem cells: from basic research to clinical application. American Journal of Cancer Research 5(5): 1602–1609.
Xu, H.D. and Qin, Z.H., 2019. Beclin 1, Bcl-2 and autophagy. Advances in Experimental Medicine and Biology 1206: 109–126. 10.1007/978-981-15-0602-4_5
Yang, Y., Chen, D., Liu, H. and Yang, K., 2019b. Increased expression of lncRNA CASC9 promotes tumor progression by suppressing autophagy-mediated cell apoptosis via the AKT/mTOR pathway in oral squamous cell carcinoma. Cell Death and Disease 10: 41. 10.1038/s41419-018-1280-8
Yang, W.E., Ho, Y.C., Tang, C.M., Hsieh, Y.S., Chen, P.N., Lai, C.T., et al. 2019a. Duchesnea indica extract attenuates oral cancer cells metastatic potential through the inhibition of the matrix metalloproteinase-2 activity by down-regulating the MEK/ERK pathway. Phytomedicine 63: 152960. 10.1016/j.phymed.2019.152960
Yap, K.M., Sekar, M., Seow, L.J., Gan, S.H., Bonam, S.R., Rani, N.N.I.M., et al. 2021. Mangifera indica (Mango): a promising medicinal plant for breast cancer therapy and understanding its potential mechanisms of action. Breast Cancer: Targets and Therapy 13(13): 471–503. 10.2147/BCTT.S316667
Zhang, Y., Tang, K., Chen, L., Du, M. and Qu, Z., 2020. Exosomal CircGDI2 suppresses oral squamous cell carcinoma progression through the regulation of MiR-424-5p/SCAI axis.Cancer Management and Research 12: 7501–7514. 10.2147/CMAR.S255687
Zhong, J.S., Huang, Y.Y., Zhang, T.H., Liu, Y.P., Ding, W.J., Wu, X.F., Xie, Z.Y., Luo, H.B. and Wan, J.Z., 2015. Natural phosphodiesterase-4 inhibitors from the leaf skin of Aloe barbadensis Miller. Fitoterapia 100: 68–74. 10.1016/j.fitote.2014.11.018
Birari, L.A., Mahajan, U.B., Patil, K.R., Patil, D.D., Bagul, N.A., Belemkar, S., et al. 2020. Aloin protects against arsenic trioxide-induced myocardial membrane damage and release of inflammatory cytokines. Naunyn Schmiedebergs Arch Pharmacology 393: 1365–1372. 10.1007/s00210-020-01833-1
Boudreau, M.D., Olson, G.R., Tryndyak, V.P., Bryant, M.S., Felton, R.P. and Beland, F.A., 2017. From the cover: aloin, a component of the aloe vera plant leaf, induces pathological changes and modulates the composition of microbiota in the large intestines of F344/N male rats. Toxicological Sciences 158: 302–318. 10.1093/toxsci/kfx105
Chang, R., Zhou, R., Qi, X., Wang, J., Wu, F., Yang, W., et al. 2016. Protective effects of aloin on oxygen and glucose deprivation-induced injury in PC12 cells. Brain Research Bulletin (BRB) 121: 75–83. 10.1016/j.brainresbull.2016.01.001
D’Arcy, M.S., 2019. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biology International 43: 582–592. 10.1002/cbin.11137
Fridman, E., Na’ara, S., Agarwal, J., Amit, M., Bachar, G., Villaret A.B., et al. 2018. The role of adjuvant treatment in early-stage oral cavity squamous cell carcinoma: an international collaborative study. Cancer 124: 2948–2955. 10.1002/cncr.31531
Ichimura, Y., Kominami, E., Tanaka, K. and Komatsu, M., 2008. Selective turnover of p62/A170/SQSTM1 by autophagy. Autophagy 4: 1063–1066. 10.4161/auto.6826
Irani, S., 2016. Distant metastasis from oral cancer: a review and molecular biologic aspects. Journal of International Society of Preventive and Community Dentistry 6: 265–271. 10.4103/2231-0762.186805
Islam, M.A., Sooro, M.A. and Zhang, P., 2018. Autophagic regulation of p62 is critical for cancer therapy. International Journal of Molecular Sciences 19: 1405. 10.3390/ijms19051405
Johnson, D.E., Burtness, B., Leemans, C.R., Lui, V.W.Y., Bauman, J.E. and Grandis, J.R., 2020. Head and neck squamous cell carcinoma. Nature Reviews Disease Primers 6: 92. 10.1038/s41572-020-00224-3
Khan, T., Ali, M., Khan, A., Nisar, P., Jan, S.A., Afridi, S., et al. 2019. Anticancer plants: a review of the active phytochemicals, applications in animal models, and regulatory aspects. Biomolecules 10(1). 10.3390/biom10010047
Khan, T., Relitti, N., Brindisi, M., Magnano, S., Zisterer, D., Gemma, S., et al. 2020. Autophagy modulators for the treatment of oral and esophageal squamous cell carcinomas. Medicinal Research Reviews 40: 1002–1060. 10.1002/med.21646
Kim, Y.C. and Guan, K.L., 2015. mTOR: a pharmacologic target for autophagy regulation. Journal of Clinical Investigation 125: 25–32. 10.1172/JCI73939
Kinarivala, N., Patel, R., Boustany, R.M., Al-Ahmad, A. and Trippier, P.C., 2017. Discovery of aromatic carbamates that confer neuroprotective activity by enhancing autophagy and inducing the anti-apoptotic protein B-cell lymphoma 2 (Bcl-2). Journal of Medicinal Chemistry 60: 9739–9756. 10.1021/acs.jmedchem.7b01199
Kitamura, N., Sento, S., Yoshizawa, Y., Sasabe, E., Kudo, Y. and Yamamoto, T., 2020. Current trends and future prospects of molecular targeted therapy in head and neck squamous cell carcinoma. International Journal of Molecular Sciences 22: 240. 10.3390/ijms22010240
Lee, I.C. and Bae, J.S., 2021. Inhibitory effects of aloin on TGFBIp-mediated septic responses. Journal of Asian Natural Products Research 23: 189–203. 10.1080/10286020.2019.1711066
Lee, W., Yang, S., Lee, C., Park, E.K., Kim, K.M., Ku, S.K., et al. 2019. Aloin reduces inflammatory gene iNOS via inhibition activity and p-STAT-1 and NF-kappaB. Food and Chemical Toxicology 126: 67–71. 10.1016/j.fct.2019.02.025
Lei, J., Shen, Y., Xv, G., Di, Z., Li, Y. and Li, G., 2020. Aloin suppresses lipopolysaccharide-induced acute lung injury by inhibiting NLRP3/NF-kappaB via activation of SIRT1 in mice. Immunopharmacology and Immunotoxicology 42: 306–313. 10.1080/08923973.2020.1765373
Li, P., Ren, K., yin Liang, Y., kai Liu J., wen Liang, Z., et al. 2020. Aloin promotes cell apoptosis by targeting HMGB1-TLR4-ERK axis in human melanoma cells. EXCLI Journal 19: 641.
Liang, L., Luo, H., He, Q., You, Y., Fan, Y. and Liang, J., 2018. Investigation of cancer-associated fibroblasts and p62 expression in oral cancer before and after chemotherapy. Journal of Cranio-Maxillofacial Surgery 46: 605–610. 10.1016/j.jcms.2017.12.016
Lin, C.W., Chin, H.K., Lee, S.L., Chiu, C.F., Chung, J.G., Lin, Z.Y., et al. 2019. Ursolic acid induces apoptosis and autophagy in oral cancer cells. Environmental Toxicology 34: 983–991. 10.1002/tox.22769
Maiuri, M.C., Zalckvar, E., Kimchi, A. and Kroemer, G., 2007. Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nature Reviews in Molecular Cell Biology 8: 741–752. 10.1038/nrm2239
McMullen, P. and McMullen, A., 1989. Ambulatory centers can be a source of liability, too. Nursing Connections 2(3): 42–44.
Quadri, M.F.A., Tadakamadla, S.K. and John, T., 2019. Smokeless tobacco and oral cancer in the Middle East and North Africa: a systematic review and meta-analysis. Tobacco Induced Diseases 17: 56. 10.18332/tid/110259
Sanchez, M., Gonzalez-Burgos, E., Iglesias, I. and Gomez-Serranillos, M.P., 2020. Pharmacological update properties of aloe vera and its major active constituents. Molecules 25: 1324. 10.3390/molecules25061324
Sharma, A., Kim, J.W. and Paeng, J.Y., 2018. Clinical analysis of neck node metastasis in oral cavity cancer. Journal of the Korean Association of Oral and Maxillofacial Surgeons 44: 282–288. 10.5125/jkaoms.2018.44.6.282
Sun, R., Zhai, R., Ma, C. and Miao, W., 2020. Combination of aloin and metformin enhances the antitumor effect by inhibiting the growth and invasion and inducing apoptosis and autophagy in hepatocellular carcinoma through PI3K/AKT/mTOR pathway. Cancer Medicine 9: 1141–1151. 10.1002/cam4.2723
Tandon, P., Dadhich, A., Saluja, H., Bawane, S. and Sachdeva, S., 2017. The prevalence of squamous cell carcinoma in different sites of oral cavity at our rural health care centre in Loni, Maharashtra—a retrospective 10-year study. Contemporary Oncology (Poznan, Poland) 21: 178–183. 10.5114/wo.2017.68628
Tao, H., Tang, T., Wang, S., Wang, Z., Ma, Y., Cai, T., et al. 2019. The molecular mechanisms of aloin induce gastric cancer cells apoptosis by targeting high mobility group box 1. Drug Design, Development and Therapy 13: 1221. 10.2147/DDDT.S201818
Torre, L.A., Bray, F., Siegel, R.L., Ferlay, J., Lortet-Tieulent, J. and Jemal, A., 2015. Global cancer statistics, 2012. Cancer Journal for Clinicians 65: 87–108. 10.3322/caac.21262
Wang, Z., Tang, T., Wang, S., Cai, T., Tao, H., Zhang, Q., et al. 2020b. Aloin inhibits the proliferation and migration of gastric cancer cells by regulating NOX2-ROS-mediated pro-survival signal pathways. Drug Design, Development and Therapy 14: 145–155. 10.2147/DDDT.S219247
Wang, J., Wang, S., Wang, Y., Wang, L., Xia, Q., Tian, et al. 2020a. Chemopreventive effect of modified zengshengping on oral cancer in a hamster model and assessment of its effect on liver. Journal of Ethnopharmacology 255: 112774. 10.1016/j.jep.2020.112774
Wei, M., Wu, Y., Liu, H. and Xie, C., 2020. Genipin induces autophagy and suppresses cell growth of oral squamous cell carcinoma via PI3K/AKT/MTOR pathway. Drug Design, Development and Therapy 14: 395–405. 10.2147/DDDT.S222694
White, E., 2012. Deconvoluting the context-dependent role for autophagy in cancer. Nature Reviews Cancer 12: 401–410. 10.1038/nrc3262
Xia, P. and Xu, X.Y., 2015. PI3K/Akt/mTOR signaling pathway in cancer stem cells: from basic research to clinical application. American Journal of Cancer Research 5(5): 1602–1609.
Xu, H.D. and Qin, Z.H., 2019. Beclin 1, Bcl-2 and autophagy. Advances in Experimental Medicine and Biology 1206: 109–126. 10.1007/978-981-15-0602-4_5
Yang, Y., Chen, D., Liu, H. and Yang, K., 2019b. Increased expression of lncRNA CASC9 promotes tumor progression by suppressing autophagy-mediated cell apoptosis via the AKT/mTOR pathway in oral squamous cell carcinoma. Cell Death and Disease 10: 41. 10.1038/s41419-018-1280-8
Yang, W.E., Ho, Y.C., Tang, C.M., Hsieh, Y.S., Chen, P.N., Lai, C.T., et al. 2019a. Duchesnea indica extract attenuates oral cancer cells metastatic potential through the inhibition of the matrix metalloproteinase-2 activity by down-regulating the MEK/ERK pathway. Phytomedicine 63: 152960. 10.1016/j.phymed.2019.152960
Yap, K.M., Sekar, M., Seow, L.J., Gan, S.H., Bonam, S.R., Rani, N.N.I.M., et al. 2021. Mangifera indica (Mango): a promising medicinal plant for breast cancer therapy and understanding its potential mechanisms of action. Breast Cancer: Targets and Therapy 13(13): 471–503. 10.2147/BCTT.S316667
Zhang, Y., Tang, K., Chen, L., Du, M. and Qu, Z., 2020. Exosomal CircGDI2 suppresses oral squamous cell carcinoma progression through the regulation of MiR-424-5p/SCAI axis.Cancer Management and Research 12: 7501–7514. 10.2147/CMAR.S255687
Zhong, J.S., Huang, Y.Y., Zhang, T.H., Liu, Y.P., Ding, W.J., Wu, X.F., Xie, Z.Y., Luo, H.B. and Wan, J.Z., 2015. Natural phosphodiesterase-4 inhibitors from the leaf skin of Aloe barbadensis Miller. Fitoterapia 100: 68–74. 10.1016/j.fitote.2014.11.018
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