Signal Transducer and Activator of Transcription 3 (STAT3), a common oncogenic mediator, is constitutively activated in many types of human cancers; therefore it is a major focus in the development of novel anti-cancer agents. Hedyotis diffusa Willd has been used as a major component in several Chinese medicine formulas for the clinical treatment of colorectal cancer (CRC). However, the precise mechanism of its anti-tumor activity remains largely unclear. Using a CRC mouse xenograft model, in the present study we evaluated the effect of the ethanol extract of Hedyotis diffusa Willd (EEHDW) on tumor growth in vivo and investigated the underlying molecular mechanisms. We found that EEHDW reduced tumor volume and tumor weight, but had no effect on body weight gain in CRC mice, demonstrating that EEHDW can inhibit CRC growth in vivo without apparent adverse effect. In addition, EEHDW treatment suppressed STAT3 phosphorylation in tumor tissues, which in turn resulted in the promotion of cancer cell apoptosis and inhibition of proliferation. Moreover, EEHDW treatment altered the expression pattern of several important target genes of the STAT3 signaling pathway, i.e., decreased expression of Cyclin D1, CDK4 and Bcl-2 as well as up-regulated p21 and Bax. These results suggest that suppression of the STAT3 pathway might be one of the mechanisms by which EEHDW treats colorectal cancer.

Qingjie Fuzheng granules (QFG) exert an anticancer effect against colorectal cancers (CRC). However, the pharmacological molecular mechanisms are still unclear. This study was aimed to establish a simple method to predict targets of QFG against CRC by the network pharmacology strategy. 461 compounds and 1559 targets in QFG were enriched by BATMAN-TCM. 21 of the common targets were obtained by the groups of “Jun,” “Chen,” “Zuo,” and “Shi” medicine in QFG. The enrichment analyses of GO functional terms, KEGG pathway, and OMIM/TTD diseases displayed the targets in the different and complementary effects of four functional medicines in QFG. Then, 613 differential targets for QFG in CRC were identified. GO functional terms and KEGG pathway analyses showed that QFG regulated the inflammatory function and lipid metabolic process. There were also targets that played a role in the binding to the receptors in membranes, in the activation of the transportation signal, and provided pain relief by regulation of the neural related pathways. Next, the protein-protein interaction network was analyzed, and the levels of the predicted targets in CRC primary tumor were explored, and 7 candidate targets of QFG against CRC were obtained. Furthermore, with real-time PCR and enzyme-linked immunosorbent assay (ELISA) analysis, downregulation of dopamine D2 receptor (DRD2) and interleukin-6 (IL-6), and upregulation of interleukin-10 (IL-10) were identified following the treatment of QFG. At last, the survival and prognosis of the potential targets of QFG in CRC patients were analyzed by GenomicScape, and IL-6 was suggested to be an index for the regulation of QFG in CRC. These results might elucidate the possible antitumor mechanism of QFG and highlight the candidate therapeutic targets and the application direction in clinical treatment for QFG.

Objective From the perspective of colorectal cancer stem cells，the purpose of this study was to explore the effect and its mechanism of Spica Prunellae on the proliferation and stemness of colorectal cancer cells based on Notch signaling pathway. Methods Colorectal cancer SW480 cells were cultured in vitro. When the cells were in logarithmic growth phase, they were randomly divided into control group, 0.1 mg/mL group, 0.2 mg/mL group and 0.4 mg/mL group. The control group did not receive drug intervention; the other three groups were added with 0.1 mg/mL, 0.2 mg/mL and 0.4 mg/mL Spica Prunellae solution, respectively. Meanwhile, DMSO in each group was supplemented to the same amount. After 24 hours of intervention, the subsequent experiments were performed. The cell growth morphology and density were observed by inverted microscope. The number of viable cells was detected by 0.4% Trypan blue staining and cell survival rate was calculated. The proliferation inhibition rate of colorectal cancer cells was detected using Cell Counting Kit-8 (CCK8) assay. Flow cytometry was used to detect the proportion of stem cells in colorectal cancer cells. The self-renewal ability of colorectal cancer cells was detected by sphere formation assay. Western blot method was used to detect the relative expression of c-Myc, CD44, CD133, SOX2 and Notch pathway-related proteins Notch1 and Hes1 in colorectal cancer cells. Results Compared with the control group, the proliferation and viability of colorectal cancer SW480 cells in the 0.1, 0.2, and 0.4 mg/mL groups were significantly inhibited with the increase of Spica Prunellae concentration (P<0.05). The proliferation ability of colorectal cancer SW480 cells significantly decreased with the increase of Spica Prunellae concentration (P<0.05). The proportion of cancer stem cells in colorectal cancer SW480 cells significantly decreased with the increase of Spica Prunellae concentration (P<0.05). The self-renewal ability of colorectal cancer SW480 cells was significantly inhibited with the increase of Spica Prunellae concentration (P<0.05). Compared with the control group, the relative protein expression of c-Myc, CD44, CD133, SOX2, Notch1, and Hes1 in colorectal cancer SW480 cells was significantly down-regulated with the increase of Spica Prunellae concentration in the 0.1, 0.2, and 0.4 mg/mL groups. Conclusion Spica Prunellae can inhibit the proliferation and stem cell properties of colorectal cancer cells, and inhibition of the Notch pathway activation may be one of the important mechanisms.