Genistein inhibits lung cancer cell stem-like characteristics by modulating MnSOD and FoxM1 expression

Zhimin Fu; Xiaocheng Cao; Lihua Liu; Xiaozheng Cao; Yinghong Cui; Xiang Li; Meifang Quan; Kaiqun Ren; A Chen; Chang Xu; Yebei Qiu; Xiangding Chen; Zheng Wang; Jianguo Cao

doi:10.3892/ol.2020.11802

Genistein inhibits lung cancer cell stem-like characteristics by modulating MnSOD and FoxM1 expression

Oncol Lett. 2020 Sep;20(3):2506-2515. doi: 10.3892/ol.2020.11802. Epub 2020 Jul 1.

Authors

Zhimin Fu^{1

2

3}, Xiaocheng Cao^{4

5

6}, Lihua Liu⁷, Xiaozheng Cao⁷, Yinghong Cui^{5

6}, Xiang Li^{5

6}, Meifang Quan^{5

6}, Kaiqun Ren^{5

6}, A Chen^{5

6}, Chang Xu^{5

6}, Yebei Qiu^{5

6}, Xiangding Chen³, Zheng Wang⁸, Jianguo Cao^{5

6}

Affiliations

¹ Department of Cardiothoracic Surgery, The First People's Hospital of Chenzhou, Chenzhou, Hunan 423000, P.R. China.
² Department of Cardiothoracic Surgery, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510630, P.R. China.
³ Department of Thoracic Surgery, Pingshan General Hospital of Southern Medical University, Shenzhen, Guangdong 518118, P.R. China.
⁴ Laboratory of Molecular and Statistical Genetics, Hunan Normal University, Changsha, Hunan 410081, P.R. China.
⁵ Department of Pharmaceutical Science, Medical College, Hunan Normal University, Changsha, Hunan 410013, P.R. China.
⁶ Key Laboratory of Study and Discover of Small Targeted Molecules of Hunan Province, Changsha, Hunan 410013, P.R. China.
⁷ Department of Pharmacology, Shenzhen People's Hospital 2nd Clinical Medical College of Jinan University, Shenzhen People's Hospital, Shenzhen, Guangdong 518020, P.R. China.
⁸ Department of Thoracic Surgery, The 2nd Clinical Medical College of Jinan University, Shenzhen People's Hospital, Shenzhen, Guangdong 518020, P.R. China.

Abstract

Manganese superoxide dismutase (MnSOD) promotes invasive and migratory activities by upregulating Forkhead box protein M1 (FoxM1) expression. The present study investigated whether modulation of MnSOD and FoxM1 expression was responsible for the antitumor effects of genistein on cancer stem-like cells (CSLCs) derived from non-small cell lung cancer cells (NSCLCs). Spheroids prepared from H460 or A549 cells were defined as lung cancer stem-like cells (LCSLCs) and were treated with genistein. The Cell Counting Kit-8 assay was performed to assess human lung fibroblast IMR-90 cell proliferation, as well as NSCLC H460 and A549 cell proliferation following treatment with genistein. MnSOD, FoxM1, cluster of differentiation (CD)133, CD44, BMI1 proto-oncogene, polycomb ring finger (Bmi1) and Nanog homeobox (Nanog) protein expression levels were examined via western blotting. The sphere formation assay was conducted to evaluate LCSLC self-renewal potential, and LSCLC migratory and invasive activities were analyzed using the wound healing and Transwell invasion assays, respectively. Knockdown and overexpression of MnSOD and FOXM1 via short hairpin-RNA or cDNA transfection were performed. The results indicated that genistein (80 and 100 µM) suppressed H460 and A549 cell viability compared with IMR-90 cells. Sub-cytotoxic concentrations of genistein (20 and 40 µM) inhibited sphere formation activity and decreased the protein expression levels of CD133, CD44, Bmi1 and Nanog in LCSLCs compared with the control group. Genistein also suppressed the migratory and invasive activities of LCSLCs compared with the control group. MnSOD and FoxM1 overexpression antagonized the effects of genistein (40 µM), whereas MnSOD and FoxM1 knockdown enhanced the inhibitory effects of genistein (20 µM) on CSLC characteristics of LCSLCs. Overall, the results suggested that genistein suppressed lung cancer cell CSLC characteristics by modulating MnSOD and FoxM1 expression levels.

Keywords: FoxM1; MnSOD; cancer stem cell; genistein; lung cancer.