Fuzheng Kang' ai decoction (扶正抗癌方) inhibits cell proliferation, migration and invasion by modulating mir-21-5p/human phosphatase and tensin homology deleted on chromosome ten in lung cancer cells

doi:10.19852/j.cnki.jtcm.2022.03.003

Journal of Traditional Chinese Medicine ›› 2022, Vol. 42 ›› Issue (3): 344-352.DOI: 10.19852/j.cnki.jtcm.2022.03.003

• Research Article • Previous Articles Next Articles

Fuzheng Kang' ai decoction (扶正抗癌方) inhibits cell proliferation, migration and invasion by modulating mir-21-5p/human phosphatase and tensin homology deleted on chromosome ten in lung cancer cells

LI Jinhua¹^,², LIU Chunping³, ZHAO Yueyang⁴, WU Wanyin⁵, SUN Pengtao¹^,², LI Longmei⁵, YANG Xiaobing⁵(), ZHOU Yushu⁵()

1 Department of Ultrasound, Guangdong Provincial Hospital of Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, China
2 Department of Ultrasound, Institute of Ultrasound in Musculoskeletal Sports Medicine, Guangdong Second Provincial General Hospital, Guangzhou 510317, China
3 State Key Laboratory of Dampness Syndrome of Chinese Medicine, Guangdong Provincial Hospital of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, China
4 Department of Hematology, Guangdong Provincial Hospital of Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, China
5 Department of Oncology, Clinical and Basic Research Team of TCM Prevention and Treatment of NSCLC, Guangdong Provincial Hospital of Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, China

Received:2020-12-12 Accepted:2021-02-16 Online:2022-06-15 Published:2022-05-20
Contact: YANG Xiaobing,ZHOU Yushu
About author:ZHOU Yushu, YANG Xiaobing, Department of Oncology, Guangdong Provincial Hospital of Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, China. zhouyushu819@163.com, yangxiaobing2002@126.com,Telephone: +86-20-81887233
Supported by:
China Postdoctoral Science Foundation(2020M672745);Research Project of Traditional Chinese Medicine Bureau of Guangdong Province(Ultrasound-delivery System to Deliver Solasonine to Regulate miR-141-3p and Enhance the Inhibition of Lung Cancer by Gefitinib)(20201149);National Natural Science Foundation of China of China(Exploring the Mechanism on Reversal of Gefitinib Resistance in Treating Non-Small Cell Lung Cancer by Fuzhengkangai Decoction base on miRNA-206/449a, 81974543: Research on Fuzheng kang'ai Formula Reversing the Drug Resistance to Gefitinib in Non-Small Cell Lung Cancer NSCLC via inducing Ferroptosis by Regulating Ferritin and 81803919: the Molecular Mechanism of Fuzheng Kang-ai Decoction Reversing EMT based on Crosstalk between Tumor and Its Microenvironment)(81503507)

Abstract

Abstract:

OBJECTIVE: To elucidate the potential molecular mechanism by which Fuzheng Kang’ai decoction (扶正抗癌方, FZKA) inhibits proliferation, migration, and invasion of lung cancer cells.

METHODS: Varying FZKA concentrations were used to manage lung cancer cells (A549 and PC9). We employed: cell counting kit-8 (CCK-8) and plate clone for-mation assays to examine the cell viability; flow cytometry (FCM) to analyze the cycle arrest; transwell and wound-healing assays to assess the cell invasion and migration, respectively. Further, a quantitative real-time polymerase chain reaction (qRT-PCR) assay was adopted to evaluate the miR-21-5p expression. For protein expression analysis, we employed the Western blot technique. Recombinant miR-21-5p overexpression adenovirus vector harboring GFP was constructed and transfected into A549 and PC9, after which we explored the effect of FZKA on miR-21-5p overexpression.

RESULTS: Notably, treatment with FZKA inhibited viability, clone-formation ability, invasion, and migration of lung cancer cells. Mechanistically, FZKA markedly suppressed miR-21-5p expression but elevated the human phosphatase and tensin homology deleted on chromosome ten (PTEN) protein level in both A549 and PC9 cells. Over-expression of miR-21-5p lowered PTEN protein expression. Besides, overexpressed miR-21-5p levels with adenovirus antagonized FZKA-upregulated PTEN protein expression.

CONCLUSION: The present study demonstrates how FZKA modulates cell biological behaviors, for instance, it impedes the proliferation by upregulating PTEN expression with miR-21-5p as the target. These findings unveil the potential novel molecular mechanisms from the microRNA aspect by which FZKA suppresses the growth of human lung cancer cells.

Key words: lung neoplasms, miR-21-5p, PTEN phos-phohydrolase, Fuzheng Kang’ai decoction

LI Jinhua, LIU Chunping, ZHAO Yueyang, WU Wanyin, SUN Pengtao, LI Longmei, YANG Xiaobing, ZHOU Yushu. Fuzheng Kang' ai decoction (扶正抗癌方) inhibits cell proliferation, migration and invasion by modulating mir-21-5p/human phosphatase and tensin homology deleted on chromosome ten in lung cancer cells[J]. Journal of Traditional Chinese Medicine, 2022, 42(3): 344-352.

Figures/Tables 4

Figure 1 Effect of FZKA on cell proliferation A, B: cell viability was detected by cell counting kit-8; C, D: colony-formation ability was measured by Wright-Giemsa Stain Kit; E, F: effect of FZKA on cell cycle arrest. aP < 0.01 and bP < 0.05 vs control (0 FZKA mg/mL). FZKA: Fuzheng Kang’ai decoction; OD: optical density.

Figure 2 Effect of FZKA on cell migration and invasion A,B: cell migration ability was determined by scratching con?uent; C, D: cell invasion ability was measured by transwell matrigel invasion assay, magnification, ×40. aP < 0.01 vs control (0 FZKA mg/mL). FZKA: Fuzheng Kang’ai decoction.

Figure 3 Effect of FZKA on miR-21-5p expression and its overexpression-induced cell proliferation, invasion and migratory

Figure 4 Effect of FZKA on the interaction between miR-21-5p and PTEN A, B: A549 cells were exposed to the increased concentration of FZKA for 24 h; C, D: A549 cells were transfected with equal doses of adeno-virus with miR-NC (AD-NC) or miR-21-5p mimic (AD-miR-21-5p) for 24 h before exposure of the cells to FZKA. aP < 0.05, bP < 0.01 vs 0 mg/mL FZKA or AD-NC. FZKA: Fuzheng Kang’ai decoction; PTEN: phosphatase and tensin homology deleted on chromosome ten.

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Fuzheng Kang' ai decoction (扶正抗癌方) inhibits cell proliferation, migration and invasion by modulating mir-21-5p/human phosphatase and tensin homology deleted on chromosome ten in lung cancer cells

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