8-bromo-7-methoxychrysin Reversed M2 Polarization of Tumor-associated Macrophages Induced by Liver Cancer Stem-like Cells
Authors: Duo Li
Zhenwei Song
Meifang Quan
Shuwen Sun
Jianguo Cao
Yinghong Cui
Hui Zou
Kaiqun Ren
Yu Zheng
Journal: Anti-Cancer Agents in Medicinal Chemistry (2017)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: Hepatocellular carcinoma; M2 polarization; tumor-associated macrophages; liver cancer stem-like cells; 8-bromo-7-methoxychrysin
Affiliations: Department of Pharmaceutical Science, Medical College, Hunan Normal University, Changsha, 410013,China.

Abstract:  Background: Hepatocellular carcinoma (HCC) is related to chronic liver inflammation. M2 polarization of tumor-associated macrophages (TAMs) in the tumor microenvironment promotes liver cancer stem-like cell (LCSLC) self-renewal capability and carcinogenicity. Therefore, reversing M2 polarization of TAMs could be an effective approach to cure HCC. Objective: To evaluate whether 8-bromo-7-methoxychrysin (BrMC) has an effect on M2 polarization of TAMs. Method: LCSLC and conditional medium were obtained by sphere forming assay. Identification of LCSLC were analyzed by sphere forming, wound-healing and invasion assay. TAM and effects of BrMC on it were validated by immunofluorescence staining, ELISA and griess assay. Expressions of cancer stem cell and macrophage marker were analyzed by western blotting. Results: Our results showed that BrMC significantly suppressed the expression of the M2 macrophage marker CD163. Furthermore, BrMC influenced the secretion profile of cytokines of TAMs. Mechanistically, BrMC reversed M2 polarization of TAMs due to inhibition of NF-κB activation. Conclusion: BrMC may be a potentially novel flavonoid agent that can be applied for disrupting the interaction of LCSLCs and TAMs. 

Retracted: No

PubPeer comments:
 Fig. 12.  Flow cytometry indicated that miR-495 overexpression and a low expression of NLRP3 caused suppression of cell apoptosis. ...(A), cell apoptosis in each group . ![file] Could the authors comment on the clusters of cells displaying the same properties, and present in multiple independent experiments? , 
 Here I have black-white reversed the miR-495-mimic panel and superimposed it on the NC panel, so that repeated cells cancel out. Note the disappearance of the upper-left quadrant and the amount of cancellation in the two lower quadrants. ![file] , 
 Fig. 6B.  ELISA and western blot analysis were applied to determine the miR- 495 expression and mRNA and protein expression of NLRP3, TNF-u03b1, IL-1u03b2, IL-18 and caspase-1. The beta-actin loading control seems familiar. At right, for comparison, Fig 11E from [Wang et al (2018)] .  E: protein bands of each factor in ASMCs of asthma mice in each group detected by western blot analysis . ![file] Despite the stylistic similarities in the WBs, there are no authors in common. , 
 Fig 11A.  Flow cytometry validated that the overexpression of miR-495 or silenced NLRP4 induced changes in cell cycle distribution... (A), the cycle distribution in each group . Should histograms overhang like this? ![file] , 
 Fig. 9A.  Western blot analysis demonstrated that miR-495 negatively correlated with the protein expression of NLRP3, TNF-u03b1, IL-1u03b2, IL-18 and caspase-1, and the opposite was found for NLRP3. ... (A), western blotting map . ![file] Other examples of this loading control: Fig 3A from [Gong et al (2018)] .  SHC3 silencing regulates the PI3K-AKTFoxO signaling pathway in vivo. Notes: Panels A-B, Relevant protein level determination by western blot analysis indicates that shRNA reduces protein levels of two isoforms, p63SHC3 and p52SHC3, simultaneously . ![file] *[left]* Fig 3B.  NKD2 mediates WNT u03b2-catenin signaling pathway in HCC cells ... B C, expression of NKD2, wnt-5a and u03b2-catenin in different transfection of HepG2 cell using western blot analysis compared with cells transfected with si-NC . *[right]* Fig. 3B from [Jin et al (2017)] .  The mRNA and protein expressions of CyclinD1, PCNA, Bcl-2 and Bax in MG-63 cells after transfection for 48 h in the blank, siRNA-NC and siRNA-DNA-PKcs groups. Note: A, mRNA expressions of CyclinD1, PCNA, Bcl-2 and Bax in MG-63 cells after transfection for 48 h; B, protein bands of CyclinD1, PCNA, Bcl-2 and Bax in the blank, siRNA-NC and siRNA-DNA-PKcs groups by Western blotting . ![] Could the authors comment on the re-use of the loading control? Is it possible to see the original gels? , 
 #5 Another example of this distinctive loading control, from another team of authors. FIGURE 8 from [Liu et al (2018)] .  Expressions of IGF-1, pPI3K, pAKT, pCREB, Cyt-c, and caspase-3 in astrocytes among the eight groups. (a) Western blot images of expressions of IGF-1, pPI3K, pAKT, pCREB, Cyt-c, and caspase-3 in astrocytes among each group . ![file] ,