Shikonin inhibits the Warburg effect, cell proliferation, invasion and migration by downregulating PFKFB2 expression in lung cancer

Abstract

Lung cancer is one of the most lethal diseases and therefore poses a significant threat to human health. The Warburg effect, which is the observation that cancer cells predominately produce energy through glycolysis, even under aerobic conditions, is a hallmark of cancer. 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 2 (PFKFB) is an important regulator of glycolysis. Shikonin is a Traditional Chinese herbal medicine, which has been reported to exert antitumor effects. The present study aimed to investigate the anticancer activity of shikonin in lung cancer. Cell Counting Kit-8 (CCK-8) and colony formation assays were used to analyze proliferation in A549 and H446 cells. Wound healing and Transwell assays were used to measure migration and invasion in A549 and H446 cells. Cell apoptosis was analyzed using flow cytometry. Lactate levels, glucose uptake and cellular ATP levels were measured using their corresponding commercial kits. Western blotting was performed to analyze the protein expression levels of key enzymes involved in aerobic glucose metabolism. Reverse transcription-quantitative PCR was used to analyze the mRNA expression levels of PFKFB2. The results of the present study revealed that PFKFB2 expression levels were significantly upregulated in NSCLC tissues. Shikonin treatment decreased the proliferation, migration, invasion, glucose uptake, lactate levels, ATP levels and PFKFB2 expression levels and increased apoptosis in lung cancer cells in a dose-dependent manner. The overexpression of PFKFB2 increased the proliferation, migration, glucose uptake, lactate levels and ATP levels in lung cancer cells, while the knockdown of PFKFB2 expression exerted the opposite effects. Moreover, there were no significant differences in lung cancer cell migration, apoptosis, glucose uptake, lactate levels and ATP levels between cells with knocked down PFKFB2 expression or treated with shikonin and the knockdown of PFKFB2 in cells treated with shikonin. In conclusion, the results of the present study revealed that shikonin inhibited the Warburg effect and exerted antitumor activity in lung cancer cells, which was associated with the downregulation of PFKFB2 expression.

Shikonin derivatives cause apoptosis and cell cycle arrest in human chondrosarcoma cells via death receptors and MAPK regulation

“…are traditionally used in Chinese medicine to treat, for example, infections, and inflammatory diseases, as well as hemorrhagic diseases and contain naphthoquinone derivatives, such as shikonin and derivatives thereof. Shikonin, the most widely studied naphthoquinone derivative, has demonstrated potent anti-cancer activity in various types of cancer cells [7][8][9][10][11]. Known mechanisms of action are the inhibition of cell proliferation, induction of apoptosis, and reduction of cell migration and invasion potential through a variety of molecular signal transduction pathways [12,13].…”

Circular RNA Sec61 subunit alpha isoform 1 by competitive absorption of microRNA‐513a‐5p mediates peroxisomal biogenesis factor 5 expression and promotes the malignant phenotype of non‐small cell lung cancer

“…Tumor cells mainly obtain energy through glycolysis to achieve rapid proliferation and metastasis, which is an important theoretical basis for tumor diagnosis at present 24,25 . Inhibition of glycolysis promotes apoptosis and inhibits tumor proliferation and metastasis 26 . Importantly, substantial evidence suggests that circRNAs can regulate glycolysis in cancer cells, such as circ_0008797 27 and circ‐MEMO1 12 .…”

Targeting glycolysis in non-small cell lung cancer: Promises and challenges

“…The combination of shikonin with cisplatin synergistically inhibited PKM2 and GLUT1, according to data from immune-histological assays of tumor tissues. Furthermore, shikonin affected the glycolysis regulator PFKFB2 at the transcriptome level in A549 and H446 cells (Sha et al, 2021;Dai et al, 2022). Alpha-hederin is derived from the Pulsatilla chinensis (Bunge) Regel (Ranunculaceae).…”