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

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. 6­phosphofructo­2­kinase/fructose­2,6­biphosphatase 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 Kit­8 (CCK­8) 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 transcription­quantitative 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 dose­dependent 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.

Endoplasmic reticulum stress triggered by Soyasapogenol B promotes apoptosis and autophagy in colorectal cancer.

AIM: Colorectal cancer (CRC) is a common human malignancy which accounts for 600,000 deaths annually at the global level. Soyasapogenol B (Soy B), an ingredient of soybean, has been found to exert anti-proliferative activities in vitro in human breast cancer cells. The current study aimed to evaluate the efficacy of Soy B in suppressing CRC. METHODS AND MATERIALS: The effect of Soy B on cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay. The effect of Soy B on cell proliferation was determined using colony formation assay. The percentage of apoptotic cells was determined by the TUNEL assay and flow cytometry following Annexin V-FITC/Propidium Iodide (PI) double staining. JC-1 staining was performed to examine the change in mitochondrial membrane potential. Autophagy was examined by acridine orange staining and mRFP-GFP-LC3 adenovirus transfection. Caspase-12 activities were determined by ELISA kit. Western blotting was used to determine the expression of relevant proteins. To investigate the role of autophagy in the pro-death and pro-apoptotic activities of Soy B, autophagy inhibitors Bafilomycin A1 (Baf-A1) and Atg5 siRNA were utilized. TUDCA and CHOP shRNA were utilized to block ER stress. Moreover, a CRC xenograft murine model was used to analyze the therapeutic efficacy of Soy B in vivo. KEY FINDINGS: Soy B treatment decreased the number of viable cells and colonies formed in CRC cell lines. Moreover, Soy B treatment promoted the apoptotic cell death via the intrinsic pathway and autophagy which positively contributed to cell death and apoptosis. In addition, our results showed that ER stress, triggered by Soy B, mediated apoptosis and autophagy. In vivo results revealed that Soy B could suppress tumor growth, which was associated with increased ER stress, accompanied with apoptosis and autophagy induction. SIGNIFICANCE: Soy B was able to promote cell death in vitro and in vivo. Our findings highlight the possibility of utilizing Soy B as a chemotherapeutic agent to prevent and treat CRC.

The Efficacy of Sodium Aescinate on Cutaneous Wound Healing in Diabetic Rats.

This study is aimed to evaluate the potential effects of sodium aescinate (SA, the sodium salt of aescin) on wound healing in streptozotocin-induced diabetic rats. An excision skin wound was created in diabetic rats, and the wounded rats were divided into three groups: I) control group, II) gel-treated group, and III) SA-treated group. The control group wounds received topically normal saline once daily for 19 days. The gel-treated and SA-treated wounds received topically 400 µl of pluronic F-127 gel (25%) and 400 µl of SA (0.3%) in pluronic gel, respectively, once daily for 19 days. SA application in diabetic rats increased the wound contraction and significantly decreased the level of the inflammatory cytokine tumor necrosis factor-alpha (TNF-α) in comparison to the gel-treated group and control group. SA application in diabetic rats also resulted in a marked increase in the level of anti-inflammatory cytokine interleukin-10 (IL-10) and activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) compared to the other groups. Histopathologically, SA-treated wounds showed better granulation tissue dominated by marked fibroblast proliferation, and wounds were covered by thick regenerated epithelial layer. Additionally, the application of only pluronic gel produced some beneficial effects in some parameters in comparison to control group, but most of them were not significantly different. These findings demonstrated that SA may effectively control and improve wound healing in diabetic rats via its anti-inflammatory and antioxidant activities.