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Photomicrographs of various endpoints scored in cytokinesis-block micronucleus cytome assay. (A) A normal binucleated cell (BNC) induced by cytochalasin B. Examples of BNCs containing a micronucleus (MN, B), a nucleoplasmic bridge (NPB, C) or a nuclear bud (NB, D) are shown (indicated by arrows). A mononucleated cell (E) and a BNC (F) at early stage of apoptosis with chromatin condensation and intact cytoplasmic and nuclear boundaries as well as cells exhibiting nuclear fragmentation into smaller bodies within an intact cytoplasmic membrane. A mononucleated cell (G) and a BNC (H) at late stage of necrosis with cytoplasm loss, swelled nuclear and damaged nuclear membrane with only a partially intact nuclear structure and often with nuclear material leaking from the nuclear boundary. Giemsa-stained DNA is in red, and the cytoplasm is in blue. Bar, 10 μm.
Source publication
Homeostasis of chromosomal instability (CIN) facilitates the origin and evolution of abnormal karyotypes that are critical for the survival and proliferation of cancer cells, but excessive CIN can result in cellular toxicity. Geraniin is a multifunctional ellagitannin found in some species of Geranuim and Phyllanthus. We employed the cytokinesis-bl...
Contexts in source publication
Context 1
... effects of geraniin on CIN of NCM460, Colo320 and Colo205 were investigated by the well-estimated CBMN-Cyt assay(11). Photomicrographs of the different endpoints scored in CBMN-Cyt assay are shown in Figure 2A-D. The results showed that geraniin significantly increased the frequency of MN and NPB in Colo320 cells in a dose-and time-dependent manner (P < 0.01; Table 1). ...
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Citations
... A previous study by our group found that polyphenols may induce high levels of GIN, thereby causing cancer cells to undergo apoptosis, such as resveratrol, tea polyphenols, and geranium. This may represent one of the mechanisms underlying their anticancer activity (25)(26)(27). Can CUR and SIs also inhibit colon cancer cell growth by altering GIN, and can the combination of the two substances exert a better tumor suppressor effect? This study aimed to use human colonic epithelial cells NCM460 and human colon cancer cells SW620 as the research objects to analyze the effects of the two active polyphenols on GIN and proliferation in normal and cancer cells, in addition to providing a theoretical basis for their application in anticancer therapy and health maintenance. ...
Curcumin (CUR) and soy isoflavones (SIs) are two plant-based polyphenols that have attracted much attention, because of their extensive anticancer and health maintenance effects. However, the relevant molecular mechanisms are still uncertain. Genomic instability (GIN) refers to a combination of gene abnormal amplification, sequence deletion, ectopic, and other types of gene damage in cells, and it is one of the main factors causing cells to lose normal physiological functions. Therefore, we used the cytokinesis-block micronucleus cytome (CBMN-Cyt) assay as the main research method to analyze the effects of CUR and SIs on the GIN of human normal colon cells NCM460 and colon cancer cells SW620. Results show that CUR (12.5 μM) could reduce the apoptosis of NCM460 and maintain its genomic stability while inhibiting the proliferation of SW620 and promoting its apoptosis. There was no difference in the promoting effect of GIN between SW620 and NCM460 using SIs (3.125-50 μM). When the two polyphenols (v/v = 1/1, 1.5625-6.25 μM) were mixed, they could promote the proliferation and GIN of the NCM460 and SW620 cells, but we did not find that combining the two produced a better effect on the cells. In conclusion, CUR has more prominent health and anticancer effects, and it may become a dietary recommendation for daily health maintenance and a potential adjuvant drug for cancer treatment.
... According to the research that was conducted, cancer cell lines colo205 and colo320 were given geraniin at concentrations of 25, 50, and 100 μg/ml over a period of 24-48 h, and the results showed that apoptosis was induced in the cells. It also promoted chromosomal instability, which helped to limit cancer cell line proliferation [192]. ...
Colon cancer affects both men and women and is the world's second most significant cause of cancer-related mortality. Colon cancer death rates have risen worldwide due to the current food habit and lifestyle, which include a lot of meat, alcohol, and not enough physical exercise. As a result, novel, less harmful pharmacological treatments for colon cancer are needed now more than ever before. Colorectal cancer (CRC) affects a significant portion of the world's population. Chemotherapy's limits, as demonstrated by side effects and resistance in CRC patients, are now being sought after despite recent breakthroughs that have improved patient care and survival. Numerous chemical compounds present in medicinal herbs have shown anti-tumor and anti-apoptotic properties against various cancers, including CRC, in animal experiments. These chemicals, which come from several phytochemical families, activate several signaling pathways. This article discusses research on the anti-CRC benefits of many plants conducted in vitro, as well as the phytochemical components of plants that may play a role in the study. Researchers are also looking into the impact of these compounds on various pathways involved in cancer signaling. According to this review, anti-CRC compounds may be generated from medicinal plants. That's why we're looking at how natural items can help treat cancer while lowering the risk of developing it.
... Hydrolyzable and non-hydrolyzable or condensed tannins are the two types of tannins that can be found. However, they are not structurally diverse [90,91]. Tannins have catechin units flavanotannins or condensed flavonoids that characterize the tannins' [92,93]. ...
Tannins are polyphenols characterized by different molecular weights that plants are able to synthetize during their secondary metabolism. Macromolecules (proteins, structural carbohydrates and starch) can link tannins and their digestion can decrease. Tannins can be classified into two groups: hydrolysable tannins and condensed tannins. Tannins are polyphenols, which can directly or indirectly affect intake and digestion. Their ability to bind molecules and form complexes depends on the structure of polyphenols and on the macromolecule involved. Tannins have long been known to be an "anti-nutritional agent" in monogastric and poultry animals. Using good tannins' proper application protocols helped the researchers observe positive effects on the intestinal microbial ecosystem, gut health, and animal production. Plant tannins are used as an alternative to in-feed antibiotics, and many factors have been described by researchers which contribute to the variability in their efficiencies. The objective of this study was to review the literature about tannins, their effects and use in ruminant nutrition.
... A morphology-oriented assessment of nuclear morphology was performed to quantify apoptotic and necrotic cells in 500 cells per treatment. Representative images of BNCs with MN, NPB, or NB and apoptotic and necrotic cells can be found in ref. [19]. The cytostasis was determined by nuclear division index (NDI), according to the formula NDI = (M1 + 2 M2 + 3 M ≥ 3)/N, where M1, M2, and M ≥ 3 represent the number of cells with 1, 2, or ≥3 nuclei and N is the total number of viable cells scored [19]. ...
... Representative images of BNCs with MN, NPB, or NB and apoptotic and necrotic cells can be found in ref. [19]. The cytostasis was determined by nuclear division index (NDI), according to the formula NDI = (M1 + 2 M2 + 3 M ≥ 3)/N, where M1, M2, and M ≥ 3 represent the number of cells with 1, 2, or ≥3 nuclei and N is the total number of viable cells scored [19]. ...
Obese subjects have a high baseline of genotoxic stress, but the underlying mechanism is poorly understood. Given that obesity is associated with high bile acids (BA) and low folate, we aimed to determine the interactive effect of folate deficient or supplementation to the genotoxicity and cytotoxicity of BA in human colon and liver cells. NCM460 and L-02 cells were cultured in folate deficient (22.6 nM) and replete (2260 nM) RPMI 1640 medium with or without 50 μM deoxycholic acid (DCA) or lithocholic acid (LCA) for 7 days. Moreover, these cells were cultured in folate supplemented (5.65, 11.3 and 22.6 μM) and standard (2.26 μM) medium with 200 μM DCA or LCA for 7 days. Genotoxicity and cytotoxicity were measured using the cytokinesis-block micronucleus cytome assay. Our results showed that under folate-replete condition, 50 μM DCA or LCA significantly increased the rate of micronuclei in NCM460 and L-02 cells. Significantly, the micronuclei-inducing effect of 50 μM DCA or LCA was further enhanced by folate deficiency. Interestingly, folate supplementation exerted a dose-dependent manner to significantly decrease the rates of micronuclei, nucleoplasmic bridges, nuclear buds, apoptosis and necrosis induced by 200 μM DCA or LCA in NCM460 and L-02 cells. In conclusion, the genotoxicity of moderate BA (50 μM) was exacerbated by folate deficiency and folate supplementation could efficiently protect cells against the genotoxicity and cytotoxicity of high BA (200 μM).
... CIN is defined as an increase in the rate at which whole chromosomes or large parts thereof are gained or lost [11] and induces extensive cell-to-cell heterogeneity (intratumoural heterogeneity; ITH) [12]. In many cancers, CIN is associated with tumour initiation, disease progression, disease recurrence, drug-resistance and poor patient outcomes [13][14][15][16][17][18][19][20]; however, it has only recently begun to be evaluated in EOC. In 2017, Penner-Goeke et al. [21] provided the first empirical evidence that CIN is present and dynamic in serial samples isolated from the ascites (i.e. ...
Objective
High-grade serous ovarian cancer (HGSOC) is the most lethal gynaecological malignancy in women with a high level of mortality, metastatic disease, disease recurrence and multi-drug resistance. Many previous studies have focused on characterising genome instability in recurrent resistant HGSOC and while this has advanced our understanding of HGSOC, our fundamental knowledge of the mechanisms driving genome instability remains limited. Chromosome instability (CIN; an increased rate of chromosome gains and losses) is a form of genome instability that is commonly associated with recurrence and multi-drug resistance in many cancer types but has just begun to be characterised in HGSOC.
Method
To examine the relationship between CIN and HGSOC, we employed single-cell quantitative imaging microscopy approaches capable of capturing the cell-to-cell heterogeneity associated with CIN, to assess the prevalence and dynamics of CIN within individual and patient-matched HGSOC ascites and solid tumour samples.
Results
CIN occurs in 90.9% of ascites samples and 100% of solid tumours, while in-depth analyses identified statistically significant temporal dynamics within the serial ascites samples. In general, aneuploidy and CIN increase with disease progression and frequently decrease following chemotherapy treatments in responsive disease. Finally, our work identified higher levels of CIN in solid tumours relative to ascites samples isolated from the same individual, which identifies a novel difference existing between solid tumours and ascites samples.
Conclusions
Our findings provide novel insight into the relationship between CIN and HGSOC, and uncover a previously unknown relationship existing between CIN in solid tumours and metastatic disease (ascites).
... Together with the analysis of NDI, a morphology-oriented assessment of nuclear morphology was performed to quantify apoptotic and necrotic cells in five hundred cells per treatment. Representative images of BNCs with MN, NPB or NB and apoptotic and necrotic cells can be found in ref. (27) ...
... Thus, one promising strategy is to elevate the CIN rate to a level beyond the tolerable threshold. We and many other authors have shown that pharmacologically or genetically induced catastrophic CIN is incompatible with the viability of transformed cells (27,(44)(45)(46)(47). Another emerging strategy is to reduce CIN. ...
Glutamine (Gln) is a non-essential amino acid central for generating building blocks and cellular energy in tumours and rapidly proliferating non-transformed cells. However, the influence of Gln on regulating chromosomal stability of transformed and non-transformed cells remain poorly understand. We hypothesised that Gln is required for maintaining a homeostatic level of chromosomal stability. To this end, transformed cells HeLa and A375 and non-transformed cells NCM460 and HUVEC cells were intervened with varying concentrations of Gln (10, 1, 0.1 and 0.01 mM), with or without cisplatin (0.1 µg/ml), for 24 h. The cytokinesis-block micronucleus (MN) assay was used to determine chromosomal instability (CIN), the extent of which is reflected by the frequency of MN, nucleoplasmic bridge (NPB) and nuclear bud (NB). We demonstrated an unexpected decrease in the spontaneous rate of MN, but not NPB and NB, after Gln restriction in HeLa and A375 cells. Gln restriction reduced cisplatin-induced MN, but not NPB and NB, in HeLa and A375 cells. We further revealed that Gln restriction suppressed the proliferation of HeLa cells with high CIN induced by nocodazole, partially explaining why Gln restriction decreased the frequency of spontaneous and cisplatin-induced MN in transformed cells. In contrast, Gln restriction increased MN and NB, but not NPB, in NCM460 cells. In HUVEC cells, Gln restriction increased MN, NPB and NB. Meanwhile, Gln restriction sensitised NCM460 cells to cisplatin-induced genotoxicity. A similar but more pronounced pattern was observed in HUVEC cells. Collectively, these results suggest that the in vitro influences of Gln metabolism on CIN depend on cellular contexts: Transformed cells require high Gln to fine tune their CIN in an optimal rate to maximise genomic heterogeneity and fitness, whereas non-transformed cells need high Gln to prevent CIN.
... Related studies have demonstrated that geraniin could inhibit migration, invasion, and drug resistance in transforming growth factor-β1treated A549 cells [6]. Geraniin induces breast cancer cell apoptosis by regulating the ROSstimulated p38 mitogen-activated protein kinase pathway [7].In human colorectal cancer cells, by inducing chromosomal instability of mutations, geraniin could selectively arrest cell cycle [8]. In ovarian cancer cells, geraniin inhibits cell growth by inhibiting the nuclear factor-κB pathway and decreasing MCL-1 expression. ...
Purpose: The effect of geraniin on human bladder transitional carcinoma was not clear, this study was thus intended to reveal it and reveal the mechanism. Methods: T24 cells were treated with 5, 10, and 20 μM of geraniin and the viability and apoptosis of T24 cells were determined using thiazolyl blue tetrazolium bromide (MTT) assay and flow cytometry. The protein expression levels of Cyclin D1, p21, BAL-2, BAX, cleaved caspase-3 and PI3K/AKT pathway were evaluated using western blot. Results: Geraniin decreased T24 cell viability and induced T24 cell cycle arrest. The proportion of T24 cells in S phase was decreased by geraniin. Besides, geraniin promoted T24 cell apoptosis and regulated PI3K/AKT pathway. Conclusion: Geraniin appears to regulate bladder cancer cell growth by decreasing the levels of PI3K and AKT phosphorylation. Thus, this agent may be useful in the management of bladder cancer Keywords: Geraniin, T24 cells, Apoptosis, PI3K/AKT signaling
... Recently, we found that geraniin induces excessive CIN in human colorectal cancer Colo205 and Colo320 cells in vitro [Guo et al. 2018b], suggesting geraniin may undermine the efficiency of SAC in colorectal cancer cells. Assessment of SAC activity and mitotic aberrations has been standardized previously in our lab [Guo and Wang 2016;Guo et al. 2017a;Guo et al. 2017c;Guo et al. 2018a]. ...
... However, molecular mechanism underlying the antitumor effects of geraniin has not been fully evaluated. Previously, we found that geraniin induced gross CIN in human colorectal cancer cells in vitro [Guo et al. 2018b]. Therefore, we ask the question whether geraniin treatment induces SAC defects in cancer cells. ...
... Coupled with our previous results [Guo et al. 2018b], we find that geraniin is nongenotoxic in human noncancerous colon epithelial NCM460 and CCD841 cells. This finding suggests that geraniin may possess better cancer-cell selectivity and therefore fewer side effects. ...
... The NCM460 and CCD841 cell lines were selected because epidemiologic studies has identified that DM is an independent risk factor for colorectal cancer [21,22], suggesting colon cells would be vulnerable to the pathological effects of hyperglycemia and low folate concentration accompanying T2DM. Our previous studies revealed that NCM460 and CCD841 possess different baseline of GIN and different sensitivities to genotoxic stress [14,23,24]. ...
... Under certain selective pressures, like chemotherapy, cells harboring specific growth advantages, such as drug resistance, will continue to proliferate and may ultimately produce a highly aggressive or drug-resistant tumor [23]. Thus, it is not surprising that CIN is frequently associated with disease recurrence and poor patient outcomes [14,22,[24][25][26][27][28][29][30][31][32]. Paradoxically however, CIN can also be associated with more favorable outcomes, but typically only in certain cancer contexts [33,34]. ...
Chromosome instability (CIN) is defined as an increased rate of chromosome gains and losses that manifests as cell-to-cell karyotypic heterogeneity and drives cancer initiation and evolution. Current research efforts are aimed at identifying the etiological origins of CIN, establishing its roles in cancer pathogenesis, understanding its implications for patient prognosis, and developing novel therapeutics that are capable of exploiting CIN. Thus, the ability to accurately identify and evaluate CIN is critical within both research and clinical settings. Here, we provide an overview of quantitative single cell approaches that evaluate and resolve cell-to-cell heterogeneity and CIN, and discuss considerations when selecting the most appropriate approach to suit both research and clinical contexts.
