Guizhi Fuling pills inhibit the proliferation, migration and invasion of human cutaneous malignant melanoma cells by regulating the molecular axis of LncRNA TPT1-AS1 / miR-671-5p

Bo Zhang

doi:10.14715/cmb/2020.66.5.26

Issue

Vol. 66 No. 5: Issue 5

Issue Published : August 6, 2020

The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.

Guizhi Fuling pills inhibit the proliferation, migration and invasion of human cutaneous malignant melanoma cells by regulating the molecular axis of LncRNA TPT1-AS1 / miR-671-5p

https://doi.org/10.14715/cmb/2020.66.5.26

Bo Zhang

Cosmetic Laser Center, Plastic Surgery Hospital of Chinese Academic of Medical Science, Peking Union Medical College, Beijing, 100144,China

Corresponding Author(s) : Bo Zhang

sanpi373737fb@126.com

Cellular and Molecular Biology, Vol. 66 No. 5: Issue 5
Article Published : July 31, 2020

Abstract

The current research was aimed to explore the effects of Guizhi Fuling Pills on the proliferation, migration and invasion of human cutaneous malignant melanoma cells and its regulation on the molecular axis of LncRNA TPT1-AS1 / miR-671-5p. Human cutaneous malignant melanoma cells A375 were cultured in vitro and randomly divided into Con group, Guizhi Fuling pills-L group, Guizhi Fuling pills-M group, Guizhi Fuling pills-H group, Guizhi Fuling pills-H + pcDNA group, Guizhi Poria pills-H + pcDNA-TPT1-AS1 group. MTT was used to detect cell proliferation. The Transwell cell test was used to detect cell migration and invasion. qRT-PCR was used to detect the expression of TPT1-AS1 and miR-671-5p. The dual-luciferase report experiment verified the targeting relationship of TPT1-AS1, miR-671-5p. Western blot was used to detect the expression of Ki-67, PCNA, MMP-2 and MMP-9. Compared with Con group, Guizhi Fuling Pills could inhibit cell proliferation, migration and invasion (p<0.05), and also inhibit the expression of Ki-67, PCNA, MMP-2, MMP-9, TPT1-AS1 (p<0.05), promote the expression of miR-671-5p (p<0.05)., and the differences between the indexes of Guizhi Fuling Pill-L group, Guizhi Fuling Pill-M group and Guizhi Fuling Pill-H group were statistically significant (p<0.05). The dual-luciferase report experiment confirmed that TPT1-AS1 could target and bind to miR-671-5p and could regulate the expression and activity of miR-671-5p. Overexpression of TPT1-AS1 could reduce the inhibitory effect of Guizhi Fuling Pills on proliferation, migration and invasion of A375 cells. Guizhi Fuling Pill may reduce the proliferation, migration and invasion of human cutaneous malignant melanoma cells by down-regulating the expression of TPT1-AS1 and up-regulating the expression of miR-671-5p.

Keywords

Guizhi Fuling Pill LncRNA TPT1-AS1 miR-671-5p cutaneous malignant melanoma proliferation migration invasion

Zhang, B. (2020). Guizhi Fuling pills inhibit the proliferation, migration and invasion of human cutaneous malignant melanoma cells by regulating the molecular axis of LncRNA TPT1-AS1 / miR-671-5p. Cellular and Molecular Biology, 66(5), 148–154. https://doi.org/10.14715/cmb/2020.66.5.26

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References

Weimin W, Lixia W, Lei W. Regulation of STAT3 signaling pathway on cutaneous malignant melanoma cells in G1-S phase. Chin J Plast Surg 2017; 33(6): 450-452.
Zhang Q, Yan J, Huang X, Liming YU, Dong X. Clinical application of Chinese medicine combined with high dose interferon in postoperative adjuvant therapy of malignant melanoma of the skin. Chin J Med Aesthet Cosmetol 2017; 23(2):122-4.
Zhang Y, Chen W, Wang H, Pan T, Zhang Y, Li C. Upregulation of miR-519 enhances radiosensitivity of esophageal squamous cell carcinoma through targeting PI3K/AKT/mTOR signaling pathway. Cancer Chemother Pharmacol 2019; 84(6):1209-18.
Renjian J, Xiaoqin Zh. Research progress in uterine fibroids treatment by Guizhi Fuling pills. J Pract Tradit Chin Med 2018; 34(5): 625-626.
Jiang H, Huang G, Zhao N, Zhang T, Jiang M, He Y, Zhou X, Jiang X. Long non-coding RNA TPT1-AS1 promotes cell growth and metastasis in cervical cancer via acting AS a sponge for miR-324-5p. J Exp Clin Cancer Res 2018; 37(1):169.
Qiu T, Wang K, Li X, Jin J. miR 671 5p inhibits gastric cancer cell proliferation and promotes cell apoptosis by targeting URGCP. Exp Ther Med 2018; 16(6):4753-8.
Shihong J, Lan L, Yaosong W. Mechanism of Guizhi Fuling pills in inhibiting the proliferation of human breast cancer cell MCF-7. Chin J Exp Tradit Med Formulae 2018; 24(15): 132-136.
Qiao L, Junyuan H, Jian W. Effects of radices trichosanthis decoction on human malignant melanoma cells apoptosis and the activity of Caspase-3. Chin J Tradit Chin Med Pharm 2015; 30(2): 534-536.
Chen X, Gao J, Yu Y, Zhao Z, Pan Y. LncRNA FOXD3-AS1 promotes proliferation, invasion and migration of cutaneous malignant melanoma via regulating miR-325/MAP3K2. Biomed Pharmacother 2019; 120:109438.
Yaosong W, Leida X, Peixu Zh. Study on the inhibition mechanism of Guizhi Fuling Pill on proliferation of MCF-7 cells from STAT3 pathway. Lishizhen Med Mater Med Res 2018; 29(12): 94-96.
Xu Ruiqing, Cao Baoli. Research progress in tumor inhibition by Guizhi Fuling Pills. Med J Liaoning 2012; 26(6): 302-304.
Jurikova M, Danihel ć½, Polák Å , Varga I. Ki67, PCNA, and MCM proteins: Markers of proliferation in the diagnosis of breast cancer. Acta Histochem 2016; 118(5):544-52.
Li CY, Wang Q, Wang XM, Li GX, Shen S, Wei XL. Gambogic acid exhibits anti-metastatic activity on malignant melanoma mainly through inhibition of PI3K/Akt and ERK signaling pathways. Eur J Pharmacol 2019; 864:172719.
Zhang Y, Sun J, Qi Y, Wang Y, Ding Y, Wang K, Zhou Q, Wang J, Ma F, Zhang J, Guo B. Long non-coding RNA TPT1-AS1 promotes angiogenesis and metastasis of colorectal cancer through TPT1-AS1/NF90/VEGFA signaling pathway. Aging (Albany NY) 2020; 12(7):6191.
Wu W, Gao H, Li X, Zhu Y, Peng S, Yu J, Zhan G, Wang J, Liu N, Guo X. LncRNA TPT1"AS1 promotes tumorigenesis and metastasis in epithelial ovarian cancer by inducing TPT1 expression. Cancer Sci 2019;110(5):1587.
Song H, Xu Y, Xu T, Fan R, Jiang T, Cao M, Shi L, Song J. CircPIP5K1A activates KRT80 and PI3K/AKT pathway to promote gastric cancer development through sponging miR-671-5p. Biomed Pharmacother 2020; 126:109941.
Xin C, Lu S, Li Y, Zhang Y, Tian J, Zhang S, Yang S, Gao T, Xu J. miR-671-5p Inhibits Tumor Proliferation by Blocking Cell Cycle in Osteosarcoma. DNA Cell Biol 2019; 38(9):996-1004.
Beheshti Ale Agha A, Kahrizi D, Ahmadvand A, Bashiri H, Fakhri R. Development of PCR primer systems for amplification of 16S-rDNA to detect of Thiobacillus spp. Cell Mol Biol 2017; 63(11).
Ghaheri M, Kahrizi D, Yari K, Babaie A, Suthar RS, Kazemi E. A comparative evaluation of four DNA extraction protocols from whole blood sample. Cell Mol Biol; 62(3):120-124.
Dehghanian F, Kay M, Kahrizi D. A novel recombinant AzrC protein proposed by molecular docking and in silico analyses to improve azo dye's binding affinity. Gene 2015; 569(2):233-8.
Bordbar M, Darvishzadeh R, Pazhouhandeh M, Kahrizi D. An overview of genome editing methods based on endonucleases. Mod Genet J 2020; 15(2): 75-92.
Ghaheri M, Kahrizi D, Bahrami G, Mohammadi-Motlagh HR. Study of gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni under various mannitol concentrations. Mol Biol Rep 2019; 46(1): 7-16.
Eruygur N, Ucar E, Akpulat HA, Shahsavari K, Safavi SM, Kahrizi D. In vitro antioxidant assessment, screening of enzyme inhibitory activities of methanol and water extracts and gene expression in Hypericum lydium. Mol Biol Rep 2019; 46(2): 2121-9.
Esmaeili F, Ghaheri M, Kahrizi D, Mansouri M, Safavi SM, Ghorbani T, Muhammadi S, Rahmanian E, Vaziri S. Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 1-5.
Akbari F, Arminian A, Kahrizi D, Fazeli A, Ghaheri M. Effect of nitrogen sources on gene expression of Stevia rebaudiana (Bertoni) under in vitro conditions. Cell Mol Biol, 64(2): 11-16.
Ghaheri M, Adibrad E, Safavi SM, Kahrizi D, Soroush A, Muhammadi S, Ghorbani T, Sabzevari A, Ansarypour Z, Rahmanian E. Effects of life cycle and leaves location on gene expression and glycoside biosynthesis pathway in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 17-22.
Kahrizi D, Ghaheri M, Yari Z, Yari K, Bahraminejad S. Investigation of different concentrations of MS media effects on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 23-27.
Hashempoor S, Ghaheri M, Kahrizi D, Kazemi N, Muhammadi S, Safavi SM, Ghorbani T, Rahmanian E, Heshmatpanaah M. Effects of different concentrations of mannitol on gene expression in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 28-31.
Akbarabadi A, Ismaili A, Kahrizi D, Firouzabadi FN. Validation of expression stability of reference genes in response to herbicide stress in wild oat (Avena ludoviciana). Cell Mol Biol 2018; 64(4). 113-118.
Ghorbani T, Kahrizi D, Saeidi M, Arji I. Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression. Cell Mol Biol, 63(11): 32-36.
Kahrizi D, Ghari SM, Ghaheri M, Fallah F, Ghorbani T, Beheshti AA, Kazemi E, Ansarypour Z. Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol, 63(7): 107-111.
Fallah F, Nokhasi F, Ghaheri M, Kahrizi D, Beheshti Ale AA, Ghorbani T, Kazemi E, Ansarypour Z. Effect of salinity on gene expression, morphological and biochemical characteristics of Stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol 2017; 63(7): 102-106.
Fatemi SS, Yari K, Tavallaei M, Kahrizi D. Expression of recombinant Hc domain of Clostridium botulinum neurotoxin A in E. coli and its purification as a vaccine candidate against botulism. Sci J Kurdistan Univ Med Sci 2011 Aug 10;16(2):36-44.
Taheri Z, Asadzadeh Aghdaei H, Irani S, Modarressi MH, Noormohammadi Z. Clinical Correlation of miR-200c/141 Cluster DNA Methylation and miR-141 Expression with the Clinicopathological Features of Colorectal Primary Lesions/Tumors. Rep Biochem Mol Biol 2019; 8(3):208-215.
Esfandi F, Fallah H, Arsang-Jang S, Taheri M, Ghafouri-Fard S. The Expression of CCAT2, UCA1, PANDA and GHET1 Long Non-coding RNAs in Lung Cancer. Rep Biochem Mol Biol 2019; 8(1):36-41.
Azari I, Ghafouri-Fard S, Omrani MD, Arsang-Jang S, Kordi Tamandani DM, Saroone Rigi M, Rafiee S, Pouresmaeili F, Taheri M. Expression of Long Non-Coding RNAs in Placentas of Intrauterine Growth Restriction (IUGR) Pregnancies. Rep Biochem Mol Biol 2019; 8(1):25-31.

References

Weimin W, Lixia W, Lei W. Regulation of STAT3 signaling pathway on cutaneous malignant melanoma cells in G1-S phase. Chin J Plast Surg 2017; 33(6): 450-452.

Zhang Q, Yan J, Huang X, Liming YU, Dong X. Clinical application of Chinese medicine combined with high dose interferon in postoperative adjuvant therapy of malignant melanoma of the skin. Chin J Med Aesthet Cosmetol 2017; 23(2):122-4.

Zhang Y, Chen W, Wang H, Pan T, Zhang Y, Li C. Upregulation of miR-519 enhances radiosensitivity of esophageal squamous cell carcinoma through targeting PI3K/AKT/mTOR signaling pathway. Cancer Chemother Pharmacol 2019; 84(6):1209-18.

Renjian J, Xiaoqin Zh. Research progress in uterine fibroids treatment by Guizhi Fuling pills. J Pract Tradit Chin Med 2018; 34(5): 625-626.

Jiang H, Huang G, Zhao N, Zhang T, Jiang M, He Y, Zhou X, Jiang X. Long non-coding RNA TPT1-AS1 promotes cell growth and metastasis in cervical cancer via acting AS a sponge for miR-324-5p. J Exp Clin Cancer Res 2018; 37(1):169.

Qiu T, Wang K, Li X, Jin J. miR 671 5p inhibits gastric cancer cell proliferation and promotes cell apoptosis by targeting URGCP. Exp Ther Med 2018; 16(6):4753-8.

Shihong J, Lan L, Yaosong W. Mechanism of Guizhi Fuling pills in inhibiting the proliferation of human breast cancer cell MCF-7. Chin J Exp Tradit Med Formulae 2018; 24(15): 132-136.

Qiao L, Junyuan H, Jian W. Effects of radices trichosanthis decoction on human malignant melanoma cells apoptosis and the activity of Caspase-3. Chin J Tradit Chin Med Pharm 2015; 30(2): 534-536.

Chen X, Gao J, Yu Y, Zhao Z, Pan Y. LncRNA FOXD3-AS1 promotes proliferation, invasion and migration of cutaneous malignant melanoma via regulating miR-325/MAP3K2. Biomed Pharmacother 2019; 120:109438.

Yaosong W, Leida X, Peixu Zh. Study on the inhibition mechanism of Guizhi Fuling Pill on proliferation of MCF-7 cells from STAT3 pathway. Lishizhen Med Mater Med Res 2018; 29(12): 94-96.

Xu Ruiqing, Cao Baoli. Research progress in tumor inhibition by Guizhi Fuling Pills. Med J Liaoning 2012; 26(6): 302-304.

Jurikova M, Danihel ć½, Polák Å , Varga I. Ki67, PCNA, and MCM proteins: Markers of proliferation in the diagnosis of breast cancer. Acta Histochem 2016; 118(5):544-52.

Li CY, Wang Q, Wang XM, Li GX, Shen S, Wei XL. Gambogic acid exhibits anti-metastatic activity on malignant melanoma mainly through inhibition of PI3K/Akt and ERK signaling pathways. Eur J Pharmacol 2019; 864:172719.

Zhang Y, Sun J, Qi Y, Wang Y, Ding Y, Wang K, Zhou Q, Wang J, Ma F, Zhang J, Guo B. Long non-coding RNA TPT1-AS1 promotes angiogenesis and metastasis of colorectal cancer through TPT1-AS1/NF90/VEGFA signaling pathway. Aging (Albany NY) 2020; 12(7):6191.

Wu W, Gao H, Li X, Zhu Y, Peng S, Yu J, Zhan G, Wang J, Liu N, Guo X. LncRNA TPT1"AS1 promotes tumorigenesis and metastasis in epithelial ovarian cancer by inducing TPT1 expression. Cancer Sci 2019;110(5):1587.

Song H, Xu Y, Xu T, Fan R, Jiang T, Cao M, Shi L, Song J. CircPIP5K1A activates KRT80 and PI3K/AKT pathway to promote gastric cancer development through sponging miR-671-5p. Biomed Pharmacother 2020; 126:109941.

Xin C, Lu S, Li Y, Zhang Y, Tian J, Zhang S, Yang S, Gao T, Xu J. miR-671-5p Inhibits Tumor Proliferation by Blocking Cell Cycle in Osteosarcoma. DNA Cell Biol 2019; 38(9):996-1004.

Beheshti Ale Agha A, Kahrizi D, Ahmadvand A, Bashiri H, Fakhri R. Development of PCR primer systems for amplification of 16S-rDNA to detect of Thiobacillus spp. Cell Mol Biol 2017; 63(11).

Ghaheri M, Kahrizi D, Yari K, Babaie A, Suthar RS, Kazemi E. A comparative evaluation of four DNA extraction protocols from whole blood sample. Cell Mol Biol; 62(3):120-124.

Dehghanian F, Kay M, Kahrizi D. A novel recombinant AzrC protein proposed by molecular docking and in silico analyses to improve azo dye's binding affinity. Gene 2015; 569(2):233-8.

Bordbar M, Darvishzadeh R, Pazhouhandeh M, Kahrizi D. An overview of genome editing methods based on endonucleases. Mod Genet J 2020; 15(2): 75-92.

Ghaheri M, Kahrizi D, Bahrami G, Mohammadi-Motlagh HR. Study of gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni under various mannitol concentrations. Mol Biol Rep 2019; 46(1): 7-16.

Eruygur N, Ucar E, Akpulat HA, Shahsavari K, Safavi SM, Kahrizi D. In vitro antioxidant assessment, screening of enzyme inhibitory activities of methanol and water extracts and gene expression in Hypericum lydium. Mol Biol Rep 2019; 46(2): 2121-9.

Esmaeili F, Ghaheri M, Kahrizi D, Mansouri M, Safavi SM, Ghorbani T, Muhammadi S, Rahmanian E, Vaziri S. Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 1-5.

Akbari F, Arminian A, Kahrizi D, Fazeli A, Ghaheri M. Effect of nitrogen sources on gene expression of Stevia rebaudiana (Bertoni) under in vitro conditions. Cell Mol Biol, 64(2): 11-16.

Ghaheri M, Adibrad E, Safavi SM, Kahrizi D, Soroush A, Muhammadi S, Ghorbani T, Sabzevari A, Ansarypour Z, Rahmanian E. Effects of life cycle and leaves location on gene expression and glycoside biosynthesis pathway in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 17-22.

Kahrizi D, Ghaheri M, Yari Z, Yari K, Bahraminejad S. Investigation of different concentrations of MS media effects on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 23-27.

Hashempoor S, Ghaheri M, Kahrizi D, Kazemi N, Muhammadi S, Safavi SM, Ghorbani T, Rahmanian E, Heshmatpanaah M. Effects of different concentrations of mannitol on gene expression in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 28-31.

Akbarabadi A, Ismaili A, Kahrizi D, Firouzabadi FN. Validation of expression stability of reference genes in response to herbicide stress in wild oat (Avena ludoviciana). Cell Mol Biol 2018; 64(4). 113-118.

Ghorbani T, Kahrizi D, Saeidi M, Arji I. Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression. Cell Mol Biol, 63(11): 32-36.

Kahrizi D, Ghari SM, Ghaheri M, Fallah F, Ghorbani T, Beheshti AA, Kazemi E, Ansarypour Z. Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol, 63(7): 107-111.

Fallah F, Nokhasi F, Ghaheri M, Kahrizi D, Beheshti Ale AA, Ghorbani T, Kazemi E, Ansarypour Z. Effect of salinity on gene expression, morphological and biochemical characteristics of Stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol 2017; 63(7): 102-106.

Fatemi SS, Yari K, Tavallaei M, Kahrizi D. Expression of recombinant Hc domain of Clostridium botulinum neurotoxin A in E. coli and its purification as a vaccine candidate against botulism. Sci J Kurdistan Univ Med Sci 2011 Aug 10;16(2):36-44.

Taheri Z, Asadzadeh Aghdaei H, Irani S, Modarressi MH, Noormohammadi Z. Clinical Correlation of miR-200c/141 Cluster DNA Methylation and miR-141 Expression with the Clinicopathological Features of Colorectal Primary Lesions/Tumors. Rep Biochem Mol Biol 2019; 8(3):208-215.

Esfandi F, Fallah H, Arsang-Jang S, Taheri M, Ghafouri-Fard S. The Expression of CCAT2, UCA1, PANDA and GHET1 Long Non-coding RNAs in Lung Cancer. Rep Biochem Mol Biol 2019; 8(1):36-41.

Azari I, Ghafouri-Fard S, Omrani MD, Arsang-Jang S, Kordi Tamandani DM, Saroone Rigi M, Rafiee S, Pouresmaeili F, Taheri M. Expression of Long Non-Coding RNAs in Placentas of Intrauterine Growth Restriction (IUGR) Pregnancies. Rep Biochem Mol Biol 2019; 8(1):25-31.

Author biographies is not available.