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Apoptosis signaling pathway. We proposed the signaling pathway of apoptosis in Ishikawa cells after CC expo- sure. The effect of apoptosis is most likely achieved through ROS generation, which, leads to mitochondrial membrane permeabilization through the alteration of Bcl-2 family proteins ratio and triggers the intrinsic apoptosis pathway without affecting the extrinsic apoptosis pathway
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Centchroman (CC) or Ormeloxifene has been shown to induce apoptosis and cell cycle arrest in various types of cancer cells. This has, however, not been addressed for endometrial cancer cells where its (CC) mechanism of action remains unclear. This study focuses on the basis of antineoplasticity of CC by blocking the targets involved in the cell cyc...
Citations
... Redox is involved in cell differentiation, metabolism, apoptosis and proliferation of tumour cells [26][27][28][29]. Previous studies have shown oxidative stress-induced DNA damage and methylation provide growth advantages for serous EC cells by activating proto-oncogenes and silencing oncosuppressor genes, which in turn affect the occurrence and development of tumours [30][31][32]. ...
In recent studies, the tumourigenesis and development of endometrial carcinoma (EC) have been correlated significantly with redox. We aimed to develop and validate a redox-related prognostic model of patients with EC to predict the prognosis and the efficacy of immunotherapy. We downloaded gene expression profiles and clinical information of patients with EC from the Cancer Genome Atlas (TCGA) and the Gene Ontology (GO) dataset. We identified two key differentially expressed redox genes (CYBA and SMPD3) by univariate Cox regression and utilised them to calculate the risk score of all samples. Based on the median of risk scores, we composed low-and high-risk groups and performed correlation analysis with immune cell infiltration and immune checkpoints. Finally, we constructed a nomogram of the prognostic model based on clinical factors and the risk score. We verified the predictive performance using receiver operating characteristic (ROC) and calibration curves. CYBA and SMPD3 were significantly related to the prognosis of patients with EC and used to construct a risk model. There were significant differences in survival, immune cell infiltration and immune checkpoints between the low-and high-risk groups. The nomogram developed with clinical indicators and the risk scores was effective in predicting the prognosis of patients with EC. In this study, a prognostic model constructed based on two redox-related genes (CYBA and SMPD3) were proved to be independent prognostic factors of EC and associated with tumour immune microenvironment. The redox signature genes have the potential to predict the prognosis and the immunotherapy efficacy of patients with EC.
... The understanding of apoptosis has been deepened gradually, and the molecular mechanism of apoptosis has been understood more and more thoroughly. However, it is also found that this process is far from as simple as originally thought, but contains complex regulatory mechanisms [6][7][8] . It has been reported that ultrasound can inhibit proliferation and promote apoptosis by activating p38 MAPK signal transduction [9] . ...
Purpose: To investigate the effect of diagnostic ultrasound on corneal apoptosis in rats.Methods: 24 male rats were randomly divided into 4 groups: control group, 10, 20 and 30 min group. The eyeballs of rats were irradiated continuously for different time lengths by Siemens ACUSON S2000 color Doppler ultrasound diagnostic instrument. 24 hours later, the animals were killed and the corneas were taken for Tunel apoptosis detection. The apoptosis rates of corneal epithelial cells, stromal cells and endothelial cells were calculated.Results: Apoptotic cells were detected in corneal epithelial cells, stromal cells and endothelial cells of normal rats. There was no significant difference between the 10 min group and the control group (P>0.05). The apoptosis rate of 20 min and 30 min groups was significantly higher than that of the control group. With the extension of irradiation time, the apoptosis rate of corneal epithelial cells, stromal cells and endothelial cells increased.Conclusion: 20 min of rat eyeball irradiated by diagnostic ultrasound can increase the apoptosis of corneal cells, and the apoptosis is aggravated with the prolongation of ultrasound irradiation time. Keywords: Cornea; Ultrasonography; Apoptosis; Epithelial cells
... In EnCa, cell cycle is a hot research direction. Shyam et al. found that centchroman induced cell-cycle arrest in human EnCa cells [10]. Qiu et al. discovered that JQ1 inhibited tumor growth by mediating PTEN/PI3K/AKT axis in EnCa [11]. ...
Background:
Endometrial cancer (EnCa) ranks fourth in menace within women's malignant tumors. Large numbers of studies have proven that functional genes can change the process of tumors by regulating the cell cycle, thereby achieving the goal of targeted therapy.
Methods:
The transcriptional data of EnCa samples obtained from the TCGA database was analyzed. A battery of bioinformatics strategies, which included GSEA, Cox and LASSO regression analysis, establishment of a prognostic signature and a nomogram for overall survival (OS) assessment. The GEPIA and CPTAC analysis were applied to validate the dysregulation of hub genes. For mutation analysis, the "maftools" package was used.
Results:
GSEA identified that cell cycle was the most associated pathway to EnCa. Five cell cycle-related genes including HMGB3, EZH2, NOTCH2, UCK2 and ODF2 were identified as prognosis-related genes to build a prognostic signature. Based on this model, the EnCa patients could be divided into low- and high-risk groups, and patients with high-risk score exhibited poorer OS. Time-dependent ROC and Cox regression analyses revealed that the 5-gene signature could predict EnCa prognosis exactly and independently. GEPIA and CPTAC validation exhibited that these genes were notably dysregulated between EnCa and normal tissues. Lower mutation rates of PTEN, TTN, ARID1A, and etc. were found in samples with high-risk score compared with that with low-risk score. GSEA analysis suggested that the samples of the low- and high-risk groups were concentrated on various pathways, which accounted for the different oncogenic mechanisms in patients in two groups.
Conclusion:
The current research construct a 5-gene signature to evaluate prognosis of EnCa patients, which may innovative clinical application of prognostic assessment.
... Centchroman (CC), a non-steroidal oral contraceptive is a type of selective estrogen receptor modulator, having estrogen antagonist action in breast, ovary and uterus & agonistic in bone [3]. We and others have demonstrated its anticancer action against various malignancies including breast, endometrial, pancreatic and head & neck cancer [4][5][6][7][8]. ...
... MCF-7/MDA MB-231 cells (0.2 × 10 6 ) were cultured in 6-well plates, incubated with 10 μM CC & 50 μM GN separately and together for 48 h. Thereafter, staining was done according to the protocol and cells were analyzed by flow cytometry [7]. ...
... Inhibition of apoptosis is a major hallmark of cancer [21]. We and others have reported that CC inhibits breast, endometrial, pancreatic and head & neck cancer cells growth by inducing apoptosis [6,7]. Herein, our result reveal that combination of CC plus GN induced higher apoptosis compared to each drug per se, evidenced by Annexin V/PI staining (Fig. 2B). ...
Aims:
Recently, strategies of cancer treatment using combination of agents with distinct molecular mechanism(s) of action are considered more promising due to its high efficacy and reduced systemic toxicity. The study is aimed to improve the efficacy of selective estrogen receptor modulator, Centchroman (CC) by combination with the phytoestrogen Genistein (GN).
Methods:
Cytotoxicity was evaluated by Sulforhodamine B assay. Cell cycle analysis was done through flow cytometry. Further, Apoptosis was analyzed using Annexin V/PI staining, tunel assay and electron microscopic examination and verified using western blot analysis. In order to validate the in vitro results, in vivo analysis was performed using 4T1-syngeneic mouse model.
Key findings:
In this study, we report that the dietary isoflavone genistein (GN) synergistically improved antineoplasticity of CC in breast cancer by arresting cells at G2/M phase culminating in ROS dependent apoptosis. The combination of CC plus GN caused dysregulation of Bax and Bcl-2 ratio inducing mitochondrial dysfunction, activation of Caspase-3/7, -9 and PARP cleavage. Further, combination significantly suppresses phosphorylation of PI3K/Akt/NF-κB, enhancing apoptosis. Additionally, combination markedly reduced tumor growth compared to CC and GN alone in mouse 4T1 breast tumor model.
Significance:
Together, these studies suggest that GN represents a potential adjunct molecule whose role in CC induced apoptosis deserves attention.
... The inhibition of cell growth could be mediated by cell cycle arrest [13,14]. To find out the molecular mechanisms involved in L61H10, we detected the changes in cell cycle distribution. ...
Pyroptosis is a novel manner of cell death that can be mediated by chemotherapy drugs. The awareness of pyroptosis is significantly increasing in the fields of anti-tumor research and chemotherapy drugs. Invoking the occurrence of pyroptosis is an attractive prospect for the treatment of lung cancer. Here, the compound L61H10 was obtained as a thiopyran derivative to compare its activity with curcumin. It was indicated that L61H10 exhibited good anti-tumor activity both in vitro and in vivo via the switch of apoptosis-to-pyroptosis, which was associated with the NF-κB signaling pathway. In addition, L61H10 had no obvious side effects both in vitro and in vivo. In brief, L61H10 is shown to be a potential anti-lung cancer agent and research on its anti-tumor mechanism provides new information for chemotherapy drug research.
... After integrating the target genes of these DEMs with the DEGs, 14 overlapping genes were identified, of which three hub genes (BIRC5, CENPF, HJURP) were associated with poor prognosis and aggressive grade of patients with EC. The results of KEGG pathway analysis are noteworthy as several studies have previously demonstrated the involvement of the cell cycle in the development of EC (21,22). HTLV-1 has been identified to cause specific T cell leukemias and lymphoma (23). ...
Endometrial cancer (EC) is one of the most common gynecological cancer types worldwide. However, to the best of our knowledge, its underlying mechanisms remain unknown. The current study downloaded three mRNA and microRNA (miRNA) datasets of EC and normal tissue samples, GSE17025, GSE63678 and GSE35794, from the Gene Expression Omnibus to identify differentially expressed genes (DEGs) and miRNAs (DEMs) in EC tumor tissues. The DEGs and DEMs were then validated using data from The Cancer Genome Atlas and subjected to gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis. STRING and Cytoscape were used to construct a protein-protein interaction network and the prognostic effects of the hub genes were analyzed. Finally, miRecords was used to predict DEM targets and an miRNA-gene network was constructed. A total of 160 DEGs were identified, of which 51 genes were highly expressed and 100 DEGs were discovered from the PPI network. Three overlapping genes between the DEGs and the DEM targets, BIRC5, CENPF and HJURP, were associated with significantly worse overall survival of patients with EC. A number of DEGs were enriched in cell cycle, human T-lymphotropic virus infection and cancer-associated pathways. A total of 20 DEMs and 29 miRNA gene pairs were identified. In conclusion, the identified DEGs, DEMs and pathways in EC may provide new insights into understanding the underlying molecular mechanisms that facilitate EC tumorigenesis and progression.
... 41 Increased oxidative stress increased ROS generation which in turn activates the mitochondrial-related apoptotic signals by oxidizing mitochondrial membrane proteins leading to loss of mitochondrial membrane potential. 42 Our investigations had demonstrated that higher concentration of compound 4d (2 × IC 50 ) remarkably enhanced the intracellular ROS production ( Figure 5) and also decreased the mitochondrial membrane potential ( Figure 6A and 6B). We had also studied that the compound 4d upregulated the expression of cyt C which leads to release of cyt C in cytosol as evident from mitochondrial dysfunction. ...
N‐benzhydrylpiperazine and 1,3,4‐oxadiazoles are pharmacologically active scaffolds which exhibits significant inhibitory growth effects against various cancer cells, however, antiproliferation effects and the underlying mechanism for inducing apoptosis for aforementioned scaffolds addressing HeLa cancer cells remains uncertain. In this study, N‐benzhydrylpiperazine clubbed with 1,3,4‐oxadiazoles (4a–4h) were synthesized, subsequently characterized using high resolution spectroscopic techniques and eventually evaluated for their antiproliferation potential by inducing apoptosis in HeLa cancer cells. The MTT assay screening results revealed that among all, compound 4d ( N‐benzhydryl‐4‐((5‐(4‐aminophenyl)‐1,3,4‐oxadiazol‐2‐yl)methyl)piperazine) in particular, exhibited IC 50 value of 28.13 ± 0.21 μg/mL and significantly inhibited the proliferation of HeLa cancer cells in concentration‐dependent manner. The in vitro anticancer assays for treated HeLa cells resulted in alterations in the cell morphology, reduction in colony formation, and inhibition of cell migration in concentration‐dependent treatment. Furthermore, G2/M phase arrest, variations in the nuclear morphology, degradation of chromosomal DNA confirmed the ongoing apoptosis in treated HeLa cells. Increase in the expression of cytochrome C and caspase‐3 confirmed the involvement of intrinsic mitochondrial pathway regulating the cell death. Also, elevation in reactive oxygen species level and loss of mitochondrial membrane potential signified that compound 4d induced apoptosis in HeLa cells by generating the oxidative stress. Therefore, compound 4d may act as a potent chemotherapeutic agent against human cervical cancer. We are reporting for the first time the underlying mechanism involved in apoptosis induction in HeLa cancer cells for structurally similar azole conjugates.
... Our investigations provide the better understanding of the mechanism of action of compound 5e and basis for the future development of drug for cervical cancer. Apoptosis is prime target for many anticancer drugs according to the various reports [30,31]. In the present study, the changes in cell morphology reveal that compound 5e is cytotoxic and causes cell death in a concentration dependent manner (Fig. 2). ...
Piperazine scaffolds or 2-Azetidinone pharmacophores have been reported to show anti-cancer activities and
apoptosis induction in different types of cancer cells. However, the mechanistic studies involve in induction of
apoptosis addressing these two moieties for human cervical cancer cells remain uncertain. The present study
emphasizes on the anti-proliferating properties and mechanism involved in induction of apoptosis for these
structurally related azoles derivatives in HeLa cancer cells. 1-phenylpiperazine clubbed with 2-azetidione
derivatives (5a-5h) were synthesized, characterized using various spectroscopic techniques and evaluated for their in
-vitro anti-proliferative activities and induction of apoptosis. Further, we also evaluated oxidative stress generated
by these synthetic derivatives (5a-5h). Cell viability studies revealed that among all, the compound N-(3-chloro-2-
(3-nitrophenyl)-4-oxoazetidin-1-yl)-2-(4-phenylpiperazin-1-yl) acetamide 5e remarkably inhibited the growth of
HeLa cells in a concentration dependent manner having IC50 value of 29.44±1.46 µg/ml. Morphological changes,
colonies suppression and inhibition of migration clearly showed the antineoplasicity in HeLa cells treated with 5e.
Simultaneously, phosphatidylserine externalization, DNA fragmentation and cell-cycle arrest showed ongoing
apoptosis in the HeLa cancer cells induced by compound 5e in concentration dependent manner. Additionally,
generation of intracellular ROS along with the decrease in mitochondrial membrane potential supported that
compound 5e caused oxidative stress resulting in apoptosis through mitochondria mediated pathway. Elevation in
the level of cytochrome c and upregulation in expression of caspase-3 clearly indicated the involvement of the
intrinsic pathway of programmed cell death. In brief; compound 5e could serve as a promising lead for the
development of an effective antitumor agent.
... To determine the cyto-static/proliferative/toxic effect of CC/DZ separately and together in HBCCs versus non-tumorigenic MCF-10A, sulforhodamine B (SRB) assay was performed as reported earlier by us (Kaushik et al., 2016;Shyam et al., 2017). Briefly, 1 × 10 4 cells were seeded in 96-well plates and exposed with CC (1-30µM) and DZ (10-200µM) alone for further 48 h. ...
... Interestingly, combination index analysis through CompuSyn software supported our observations too (Fig 1d), similar to GN as reported earlier (Kaushik et al., 2016). Earlier, we reported that CC by itself is able to induce apoptosis in HBCCs and Human Endometrial Cancer Cells (HECCs) (Nigam et al., 2008;Shyam et al., 2017). More, we have demonstrated the improved efficacy of CC action vis-àvis apoptosis through supplementation with resveratrol & curcumin (Singh et al., 2012). ...
Background:
Despite advancements in the prognosis and management of breast cancer, it remains a major cause of mortality in women worldwide. Centchroman (CC), an oral contraceptive has been found to exhibit anti-cancer potential against a wide range of cancer including breast cancer.
Purpose:
The present study is intended to evaluate the ability of soy isoflavone Daidzein (DZ) in enhancing the efficacy of CC in Human Breast Cancer Cells (HBCCs).
Methods/study design:
Sulforhodamine B assay was employed to determine the cytotoxicity induced by 10 µM CC & 50 µM DZ separately and together in MCF-7/MDA MB-231 HBCCs and non-tumorigenic Human Mammary Epithelial Cells (HMECs) MCF-10A as a control. Combination Index (CI) analysis was executed using CompuSyn software. Further, apoptosis was assessed using Annexin V/PI, AO/PI staining and tunel assay. Cell cycle, reactive oxygen species generation and mitochondrial membrane potential alteration was determined using flow cytometry. Western blot analysis was performed to check the expression of respective proteins.
Results:
The results suggest that the combination exerts elevated toxicity as compared to control and each drug per se without affecting HMECs MCF-10A. This therefore implies cancer cell specific action of CC plus DZ administered together. Additionally, combination index analysis suggests synergistic action of CC and DZ combination in HBCCs. Cell cycle analysis, Annexin V/PI staining, tunel assay and western blot analysis confirms the induction of apoptosis by combination in HBCCs. Interestingly, western blot analysis also revealed that the combination down-regulated the expression of proteins involved in cell survival i.e. PI3K, Akt and mTOR, suggesting inhibition of cell survival pathway.
Conclusion:
The results overall demonstrate that CC plus DZ has higher anticancer efficacy as compared to either drug alone. Hence, the combination of CC plus DZ may offer a novel strategy for the management of breast cancer.
... In Head and Neck cancer, it acts through the modulation of PI3K/mTOR Pathway [8]. CC has also been found to be effective against Chronic Myeloid Leukemia and Endometrial cancer where it induces G0/G1 growth arrest and ERK-mediated apoptosis [9,10]. Recently, CC has shown to possess anti-cancerous potential against endometrial cancer, ovarian cancer, pancreatic cancer [10][11][12]. ...
... CC has also been found to be effective against Chronic Myeloid Leukemia and Endometrial cancer where it induces G0/G1 growth arrest and ERK-mediated apoptosis [9,10]. Recently, CC has shown to possess anti-cancerous potential against endometrial cancer, ovarian cancer, pancreatic cancer [10][11][12]. Though all these studies demonstrate that CC has anti-cancerous potential, however, its role in breast cancer angiogenesis remains to be elucidated. ...
Aims:
Angiogenesis is a recognized hallmark of cancer which promotes cancer cell progression and metastasis. Inhibition of angiogenesis to attenuate cancer growth is becoming desirable strategy for breast cancer management. The present study is aimed to investigate the antiangiogenic efficacy of a novel selective estrogen receptor modulator Centchroman (CC) on human breast cancer cells.
Main methods:
Effect of CC on cell viability was evaluated using Sulforhodamine B assay. Endothelial cell proliferation, wound healing, Boyden chamber cell invasion, tube formation and chorioallantoic membrane (CAM) assays were performed to assess the effect of CC on migration, invasion and angiogenesis. Apoptosis, reactive oxygen species generation, caspase-3/7 and intracellular calcium ion level were measured through flow cytometry. Expression levels of HIF-1α, VEGF, VEGFR2, AKT and ERK were assessed by western blot analysis.
Key findings:
CC selectively induces apoptosis in human breast cancer cells without affecting non-tumorigenic breast epithelial cells MCF-10A. Moreover, it inhibits migratory, invasive and mammosphere forming potential of breast cancer. Furthermore, CC also inhibited VEGF-induced migration, invasion and tube formation of HUVECs in vitro. CC effectively inhibited neovasculature formation in chicken CAM. Western blot analysis demonstrated that CC inhibited expression of HIF-1α and its downstream target VEGF. Interestingly, CC also suppressed VEGFR2 phosphorylation and consequently attenuated AKT and ERK phosphorylation.
Significance:
Our findings suggest that CC downregulates VEGF-induced angiogenesis by modulating HIF-1α/VEGFR2 pathway and recommend it (CC) as a potential therapeutic drug for breast cancer treatment.
