Verbascoside promotes apoptosis by regulating HIPK2–p53 signaling in human colorectal cancer

Abstract

BackgroundWe investigated the role of the HIPK2–p53 signaling pathway in tumorigenesis and resistance to the drug Verbascoside (VB) in colorectal cancer (CRC), using in vivo and in vitro experiments.MethodsPrimary human CRC samples and normal intestinal tissues from patients were analyzed for HIPK2 expression by immunohistochemistry (IHC) and its expression was correlated against patients’ clinicopathological characteristics. Human CRC HCT-116 cells were implanted in BALB/c nude mice; mice with xenografted tumors were randomly administrated vehicle (control), 20, 40, or 80 mg/mL VB, or 1 mg/mL fluorouracil (5-FU). HIPK2, p53, Bax, and Bcl-2 expression in these tumors were determined by IHC. In vitro effects of VB on CRC cell proliferation and apoptosis were measured by CCK-8 assay and flow cytometry; HIPK2, p53, p-p53, Bax, and Bcl-2 were measured by western blot.ResultsIHC analysis for 100 human CRC tumor samples and 20 normal intestinal tissues, showed HIPK2 expression to inversely correlate with Dukes stage and depth of invasion in CRC (P < 0.05). In vivo, the inhibition rates of 20, 40, and 80 mg/mL VB on CRC xenograft tumor weight were 42.79%, 53.90%, and 60.99%, respectively, and were accompanied by increased expression of HIPK2, p53, and Bax, and decreased Bcl-2 expression in treated tumors. In vitro, VB significantly inhibited proliferation of CRC cell lines HCT-116, HT-29, LoVo, and SW620, in a time- and dose-dependent manner. The apoptosis rates of 25, 50, and 100 μM VB on HCT-116 cells were 10.83 ± 1.28, 11.25 ± 1.54, and 20.19 ± 2.87%, and on HT-29 cells were 18.92 ± 6.12, 21.57 ± 4.05, and 25.14 ± 6.73%, respectively. In summary, VB treatment significantly enhanced the protein expression of pro-apoptotic HIPK2, p53, p-p53, Bax, and decreased anti-apoptotic Bcl-2 expression in CRC cells.ConclusionsHIPK2 protein modulates the phosphorylation status of p53, and levels of Bax and Bcl-2 in CRC. We also found that VB effectively activated the HIPK2–p53 signaling pathway, resulting in increased CRC cell apoptosis.

Verbascoside Reverses TGF- β1-Induced Renal Cellular Fibrosis

“…Reportedly, Verbascoside is regulated in different organ cells, including Verbascoside promoted apoptosis via p53 in human CRC cells. 27 Furthermore reported that Verbascoside attenuates LPS-induced pro-inflammatory mediator production via TAK-1/JNK/AP-1 signalling in U937 cells. 37 Therefore, the role of…”

“…[17][18][19][20][21] More importantly, Verbascoside also demonstrated various biological properties including anti-hepatotoxic, antinflammatory, anti-nociceptive, antioxidant and antihemolytic effects and antineoplastic properties in addition to numerous woundhealing and neuroprotective properties. 17,19,20,[22][23][24][25][26][27][28][29][30][31][32] However, literature no survey supporting the therapeutic effects of Verbascoside in treatment of renal fibrosis.…”

Development of a High-Throughput Assay for Inhibitors of the Polo-Box Domain of Polo-Like Kinase 1 Based on Time-Resolved Fluorescence Energy Transfer

“…Among the initial 39 hit compounds, five compounds such as haematoxylin, verbascoside, menadione, lithospermic acid and DITMD, showed a robust sigmoidal inhibitory dose response with an IC 50 ranging from 1.4 to 2.8 µM which were as active as purpurogallin (2.6±0.3 µM). It has been reported that haematoxylin, verbascoside, menadione and lithospermic acid possess anticancer activities such as anti-tumor action in mice, 25) promotion of apoptosis in human colorectal cancer, 26) induction of tumor tissue necrosis against pancreatic cancer, 27) and cytotoxic activity on human breast cancer cells, 28) respectively. The pharmacological information of DITMD has not yet been reported.…”

Verbascoside Inhibits Glioblastoma Cell Proliferation, Migration and Invasion While Promoting Apoptosis Through Upregulation of Protein Tyrosine Phosphatase SHP-1 and Inhibition of STAT3 Phosphorylation

“…In addition, TMZ administration adjuvant to radiotherapy might promote the elimination of glioblastoma stem cells and consequently prolong the survival of patients and has been used as first-line treatment for glioblastoma patients [8,9]. Verbascoside (VB) is a cancer therapeutic for colorectal cancer, glioma, and breast cancer [10]. Herein, TMZ was enrolled in the present study as a comparator with the function of VB.…”

“…VB is a component of Cistanche (a medical plant in traditional Chinese medical science) in the form of polyphenols, and it has high antioxidant power, which is inhibitory to cell proliferation and facilitated cell apoptosis in treating colorectal cancer via the homeodomain interacting protein kinase 2-p53 signaling pathway [10,12]. Inactivation of tumor suppressor phosphatases is known as a critical event in the pathogenesis of brain tumor glioblastoma [13].…”