The alkaloid sanguinarine is effective against multidrug resistance in human cervical cells via bimodal cell death
Introduction
Sanguinarine (Scheme 1) is derived from the plant Sanguinaria canadensis[1]. Its principal pharmacologic use to date is in dental products based on its antibacterial, antifungal, and anti-inflammatory activities, which reduce gingival inflammation and supragingival plaque formation [2], [3], [4]. Sanguinarine also has been reported to have antiviral and tumor-targeting activity [5], [6], [7].
Molecular biological studies indicate that sanguinarine has multiple cellular targets [8]. For example, it can interact with and intercalate DNA [9], [10], inhibit microtubule assembly [11], affect membrane permeability [12], [13], and inhibit a wide variety of enzymes, including Na+/K+ ATPase [14]. Most interestingly, it also is a potent inhibitor of protein kinases [15] and NF-κB [16], which are involved in signal transduction pathways leading to cell proliferation and/or cell death [7].
Cell death is important for normal homeostasis, cell proliferation, and differentiation. The importance of cell death is demonstrated by the observation that dysregulation of cell death can lead to cancer, developmental abnormalities, and autoimmune disorders [17], [18], [19]. Cells undergoing PCD (or apoptosis) are characterized by morphologic changes, including cellular shrinkage, blebbing, and nuclear DNA condensation with or without fragmentation [20], [21], [22], [23], [24]. However, it is stated that apoptosis is rarely observed in vivo and may not be the sole mechanism of cell death [25]. The discovery of intact novel forms of cell death pathways induced by potential anticancer agents may have an important bearing in overcoming chemoresistance.
Of all neoplasms found in females worldwide, cervical cancer has the third highest incidence and is fourth on the list of the leading causes of death by cancer [26], [27]. The available drugs most commonly used for treating cervical malignancies are impeded by frequent progression to chemotherapy resistance. Sanguinarine may be effective against MDR, since the related Sanguinaria canadensis-derived alkaloid, chelerythrine, has been shown to be cytotoxic to cancer cells and MDR cells [28]. In this study, we used our recently established in vitro cervical cancer model system for MDR [29] to investigate whether sanguinarine is effective against MDR in human cervical cells, and to understand the cellular and molecular mechanisms by which it may induce cell death.
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Cell culture, cell viability assays, and clonogenic survival assays
Most cell culture protocols, the HPV type 16-immortalized human endocervical cell line (HEN-16-2), the CSC-transformed HEN-16-2 cell line (HEN-16-2T), and the MDR HEN-16-2 cell line (HEN-16-2/CDDP) have been described previously [29], [30], [31]. Cells were cultured in keratinocyte growth medium (KGM). HeLa cervical carcinoma, CEM-VLB leukemia, CEM-T4 leukemia, K562 erythroleukemia, and JM1 pre-B cell lymphoblastic cells were obtained from the American Type Culture Collection. HeLa cells were
Evasion of MDR of human cervical HEN-16-2/CDDP cells by sanguinarine
We examined the chemotherapeutic potential of sanguinarine for MDR cervical cancer cells in a human cervical in vitro system, which is composed of MDR HEN-16-2/CDDP cells and their drug-sensitive parental HEN-16-2 cells [29]. Cell viability, measured by the trypan blue exclusion assay, was similar in both types of cells treated with 0, 0.13, 0.26, 0.53, 1.06, 2.12, and 4.24 μM sanguinarine for 4 or 48 hr (Table 1; Fig. 1). The propidium iodide exclusion assay also showed no significant difference
Discussion
We established the in vitro MDR cervical cell system used in this report by treating HPV16-immortalized human endocervical HEN-16-2 cells with cisplatin [29]. Cell viability was significantly higher in the MDR HEN-16-2/CDDP cells than in the parental cells after treatment with cisplatin, actinomycin D, doxorubicin, etoposide, paclitaxel, 5-fluorouracil, staurosporine, heat shock, or UV radiation [29], [39]. However, this study found no significant difference in the effect of sanguinarine on
Acknowledgements
We thank Mr. G. Chernenko, Ms. Y. Hao, and Ms. L. Lee for excellent technical assistance. The work was supported by a National Cancer Institute of Canada grant (2734 to A.P.) with funds from the Canadian Cancer Society; Medical Research Council of Canada grants (MT-9782 and MT-10140 to A.P. and MT-13178 to A.L.); and a Canadian Institutes of Health Research (CIHR) grant (ROP-40859 to A.P.).
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2022, Translational OncologyCitation Excerpt :In purpose of searching for effective pharmaceutical of SCLC, we performed high-throughput screening with a natural compound library, and identified Sanguinarine chloride (Sg) with excellent anti-SCLC activity. Sg is a natural compound derived from Macleaya cordata and recent studies have highlighted that Sg have different degrees of anti-tumor effects on prostate cancer [7,8], cervical cancer [9], breast cancer [10–12], colorectal cancer [13], gastric cancer [14], melanoma [15,16], pancreatic cancer [17] and non-small cell lung cancer (NSCLC) [18]. But the role of Sg in SCLC has not been elucidated.