4-Acetyl-Antroquinonol B Suppresses SOD2-Enhanced Cancer Stem Cell-Like Phenotypes and Chemoresistance of Colorectal Cancer Cells by Inducing hsa-miR-324 re-Expression

Abstract

Background: Colorectal cancer (CRC) remains a leading cause of cancer-related morbidity and mortality in both sexes globally. This is not unconnected with the heterogeneity and plasticity of CRC stem cells (CRC-SCs) which stealthily exploit the niche-related and (epi)genetic factors to facilitate metastasis, chemoresistance, tumor recurrence, and disease progression. Despite the accumulating evidence of the role of dysregulated microRNAs in malignancies, the therapeutic efficacy of pharmacological-targeting of CRC-SC-associated microRNAs is relatively under-explored. Experimental approach: In this present study, we employed relatively new bioinformatics approaches, analyses of microarray data, Western blot, real-time polymerase chain reaction (RT-PCR), and functional assays to show that hsa-miR-324-5p expression is significantly suppressed in CRC cells, and inversely correlates with the aberrant expression of SOD2. Results: This converse hsa-miR-324-5p/SOD2 relationship is associated with enhanced oncogenicity, which is effectively inhibited by 4-acetylantroquinonol B (4-AAQB), as evidenced by inhibited cell viability and proliferation, as well as attenuated migration, invasion, and clonogenicity in 4-AAQB-treated DLD1 and HCT116 cells. Interestingly, 4-AAQB did not affect the viability and proliferation of normal colon cells. We also showed that 4-AAQB-induced re-expression of hsa-miR-324-5p, akin to short-interfering RNA, reduced SOD2 expression, correlates with the concurrent down-regulation of SOD2, N-cadherin, vimentin, c-Myc, and BcL-xL2, with concomitant up-regulation of E-cadherin and BAX2 proteins. Enhanced expression of hsa-miR-324-5p in the CRC cells suppressed their tumorigenicity in vitro and in vivo. Additionally, 4-AAQB synergistically potentiates the FOLFOX (folinate (leucovorin), fluorouracil (5FU), and oxaliplatin) anticancer effect by eliciting the re-expression of SOD2-suppressed hsa-miR-324, and inhibiting SOD2-mediated tumorigenicity. Conclusion: Our findings highlight the pre-clinical anti-CSC efficacy of 4-AAQB, with or without FOLFOX in CRC, and suggest a potential novel therapeutic strategy for CRC patients.

LdrB Toxin with In Vitro and In Vivo Antitumor Activity as a Potential Tool for Cancer Gene Therapy

“…A spheroid model is a way to get closer to the clinical situation when studying cancer drugs and radiotherapy and, therefore, avoid the suffering of animals that, ethically, we have to minimize [33]. Moreover, these results are similar to those obtained in tumors treated with the main drugs of CRC chemotherapeutics such as Fluorouracil [34] or FOLFOX [35,36] but without their numerous side effects. Moreover, Li and collaborators described that FOLFOX caused a sharp loss of body weight accompanied by swelling and anal bleeding that have forced suspension of the treatment to allow the mice to recover after only one week of treatment [35].…”

Comprehensive Analysis of the Expression Profiles of Long Non-Coding RNAs with Associated ceRNA Network Involved in the Colon Cancer Staging and Progression

“…In addition to lncRNAs, hsa-miR-324-5p and several mRNAs including WNT11, FGFR3, PLCB4, and IKBKB were revealed to be related with the pathological stage and development of CC. Hsa-miR-324-5p is associated with enhanced oncogenicity and up-expression of hsa-miR-324-5p in the colorectal cancer cells suppresses CRC tumorigenicity in vitro as well as in vivo 25 . It might act as a therapeutic target for CC treatment.…”

4-Acetyl-Antroquinonol B Improves the Sensitization of Cetuximab on Both Kras Mutant and Wild Type Colorectal Cancer by Modulating the Expression of Ras/Raf/miR-193a-3p Signaling Axis

“…Vertical cell motility was evaluated using the matrigel in the Boyden chamber method as previously described by Justus CR, et al (2014) [44]. The tumor sphere-formation assay was performed according to our previously described method with modifications [17,18]. A small number of colon cancer cells were seeded (2000 cells/well) in six-well ultralow attachment plates (Corning, Corning, NY, USA) in serum-free media consisting of Dulbecco's modified Eagle medium (DMEM)/Ham's F12 (1:1), human epidermal growth factor (hEGF, 20 ng/mL), basic fibroblast growth factor (bFGF; 10 ng/mL (PeproTech, Rocky Hill, NJ, USA), 2 µg/mL 0.2% heparin (Sigma, St. Louis, MO, USA), and 1% penicillin/streptomycin (P/S, 100 U/mL, Hyclone, Logan, UT, USA).…”

“…In a broad spectrum of cancers, 4-acetyl-antroquinonol B (4-AAQB) isolated and purified from A. cinnamomea exerts antiproliferative effects [14][15][16]. Our previous study illustrated the anti-CRC role of 4-AAQB, which is mediated through the inhibition of the formation of CRC cancer stem cells and reactive oxygen species (ROS) oxidative stress, resulting in the modulation of CRC cells' innate or acquired insensitivity towards chemotherapy [17,18]. Growing evidence exists of the role of small non-coding RNA, particularly micro-RNA (miRNA's), in controlling the key biological process, including deciding the fate of cancer treatment [19], such as the tumor-suppressive effect of miR-193a-3p in lung cancer through targeting KRAS expression [20,21].…”