scholarly journals Evaluation of anti-cancer potential of Excoecaria agallocha (L.) leaf extract on human cervical cancer (SiHa) cell line and assessing the underlying mechanism of action

2022 ◽  
Vol 8 (1) ◽  
Author(s):  
Tamanna Sultana ◽  
Arup Kumar Mitra ◽  
Satadal Das
Keyword(s):  
Cervical Cancer ◽  
Plant Extract ◽  
Underlying Mechanism ◽  
Excoecaria Agallocha ◽  
Cycle Arrest ◽  
Siha Cells ◽  
Anti Cancer ◽  
M Phase ◽  
Extract Fraction ◽  
Stimulation Of

Abstract Background The incidence of cervical cancer is increasing at an alarming rate in many countries and presently, it is the most common form of malignant cancer being reported among women in India. Development of novel approach for cervical cancer therapy, sparing healthy normal cells overcoming the limitations of prevailing therapies is of prime importance. Mangroves constitute a significant repository of medicinally important plants. Thus, in this study, we aimed to determine the anticancer activity of the mangrove Excoecaria agallocha L. leaf extracts on human cervical cancer (SiHa HPV 16+) cell line with subsequent characterization of the bioactive compounds conferring the anticancer activity and studying the probable underlying mechanism of action of the purified plant extract. Results The plant extract was subjected to silica gel column chromatography and the fractions obtained were analyzed for cytotoxic activity against SiHa cells by MTT assay. One out of the three eluted fractions exhibited selective toxicity against SiHa cells with an IC50 value of 15.538 ± 0.577 µg/mL, while it had no cytotoxic effect on normal healthy human peripheral blood mononuclear cells. High-resolution liquid chromatography mass spectroscopy, coupled to electron spray ionization and diode array detection analysis, led to the structure elucidation and identification of a few pharmacologically important compounds, with Bergenin being present in the highest abundance. Fluorescence microscopy results revealed that the plant extract fraction induced LC3 puncta formation, in EGFP- SiHa cells indicating the onset of autophagy, with simultaneous stimulation of mitophagy. The plant extract also inhibited proliferation of the SiHa-smac-mCherry cells by second mitochondria-derived activator of caspase (SMAC)—induced cytochrome c dependent apoptosis, that was further confirmed with Caspase-3 activation by colorimetric assay. The GFP-dgn in SiHa cells was remarkably protected from proteasomal degradation that might upregulate the survivability of the cells significantly. Flow cytometry followed by Western blot analysis further asserted the ability of the plant extract fraction to cause cell cycle arrest of SiHa cells in the G2/M phase by significantly reducing protein expression levels of cyclin B1 and D1, decreasing Cdc2 level and simultaneously increasing p21 and p53 levels. Conclusion It could be inferred that the aqueous extract of E. agallocha successfully decreased the proliferation of SiHa cervical cancer cells through induction of autophagy and apoptosis in a concerted manner, with simultaneous stimulation of mitophagy and G2/M phase cell cycle arrest, hinting at Bergenin being the major compound conferring the anti-cancer activity of the plant extract. Thus, isolation of the identified bioactive compounds from E. agallocha and their subsequent purification for drug development might serve as a novel medicinal approach for the treatment of cervical cancer in conjugation with existing therapeutic methods.

scholarly journals Influence of icariin on inflammation, apoptosis, invasion, and tumor immunity in cervical cancer by reducing the TLR4/MyD88/NF-κB and Wnt/β-catenin pathways

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Chunyang Li ◽  
Shuangqing Yang ◽  
Huaqing Ma ◽  
Mengjia Ruan ◽  
Luyan Fang ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Underlying Mechanism ◽  
Tissue Growth ◽  
Migration And Invasion ◽  
Dependent Manner ◽  
Antitumor Effects ◽  
Siha Cells ◽  
Cervical Cancer Cells

Abstract Background Cervical cancer is a type of the most common gynecology tumor in women of the whole world. Accumulating data have shown that icariin (ICA), a natural compound, has anti-cancer activity in different cancers, including cervical cancer. The study aimed to reveal the antitumor effects and the possible underlying mechanism of ICA in U14 tumor-bearing mice and SiHa cells. Methods The antitumor effects of ICA were investigated in vivo and in vitro. The expression of TLR4/MyD88/NF-κB and Wnt/β-catenin signaling pathways were evaluated. Results We found that ICA significantly suppressed tumor tissue growth and SiHa cells viability in a dose-dependent manner. Also, ICA enhanced the anti-tumor humoral immunity in vivo. Moreover, ICA significantly improved the composition of the microbiota in mice models. Additionally, the results clarified that ICA significantly inhibited the migration, invasion capacity, and expression levels of TGF-β1, TNF-α, IL-6, IL-17A, IL-10 in SiHa cells. Meanwhile, ICA was revealed to promote the apoptosis of cervical cancer cells by down-regulating Ki67, survivin, Bcl-2, c-Myc, and up-regulating P16, P53, Bax levels in vivo and in vitro. For the part of mechanism exploration, we showed that ICA inhibits the inflammation, proliferation, migration, and invasion, as well as promotes apoptosis and immunity in cervical cancer through impairment of TLR4/MyD88/NF-κB and Wnt/β-catenin pathways. Conclusions Taken together, ICA could be a potential supplementary agent for cervical cancer treatment.

scholarly journals Glucocappasalin Induces G2/M-Phase Arrest, Apoptosis, and Autophagy Pathways by Targeting CDK1 and PLK1 in Cervical Carcinoma Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Guangya Xu ◽  
Xueling Yan ◽  
Zhongjia Hu ◽  
Lulu Zheng ◽  
Ke Ding ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Cervical Carcinoma ◽  
Hela Cells ◽  
Carcinoma Cells ◽  
Dependent Manner ◽  
Cycle Arrest ◽  
Phase Arrest ◽  
M Phase

Glucocappasalin (GCP), a natural product derived from the seeds of Descurainia sophia (L.) Webb. ex Prantl, exhibits potential antitumor activity in HeLa cervical carcinoma cells. In this study, we investigated the anti-cervical cancer property of GCP through the induction of cell cycle arrest, apoptosis, and autophagy in vitro and in vivo, and elucidated the underlying molecular mechanisms. We demonstrated that treatment with GCP inhibited the growth of HeLa, Siha, and Ca Ski cell lines in a dose-dependent manner, with HeLa cells displaying particular sensitivity to the GCP treatment. Subsequently, the expression of cyclin-dependent kinase 1 (CDK1) and polo like kinase 1 (PLK1) were evaluated in HeLa cells using the CDK1 kinase assay kit, the fluorescence polarization assay, real-time quantitative PCR, and western blotting. Our results demonstrate that GCP could be employed to attenuate the expression of CDK1 and PLK1 in a dose- and time-dependent manner. The complementary results obtained by flow cytometry and western blotting allowed us to postulate that GCP may exhibit its antitumor effects by inducing G2/M cell cycle arrest. Moreover, HeLa cells treated with GCP exhibited a loss in mitochondrial membrane potential, together with the activation of caspases 3 and 9, and poly ADP-ribose polymerase (PARP). Additionally, we found that GCP could increase the formation of acidic vesicular organelles (AVOs), as well as the levels of Beclin1, LC3-II, p62, and Atg5 proteins in HeLa cells. Further studies indicated that GCP triggered autophagy via the suppression of the PI3K/AKT/mTOR signaling pathways. The autophagy inhibitor 3-methyladenine (3-MA) was used to determine whether autophagy affects the apoptosis induced by GCP. Interestingly, the inhibition of autophagy attenuated apoptosis. In vivo anti-tumor experiments indicated that GCP (60 mg/kg, i.p.) markedly reduced the growth of HeLa xenografts in nude mice without apparent toxicity. Taken together, we demonstrate that GCP induces cell cycle G2/M-phase arrest, apoptosis, and autophagy by acting on the PI3K/AKT/mTOR signaling pathways in cervical carcinoma cells. Thus, GCP may represent a promising agent in the eradication of cervical cancer.

scholarly journals An in vitro approach to combat multidrug resistance in Salmonella typhi and human colon cancer with Excoecaria agallocha L. extract

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Tamanna Sultana ◽  
Arup Kumar Mitra ◽  
Satadal Das
Keyword(s):  
Colon Cancer ◽  
Cell Line ◽  
Plant Extract ◽  
Human Colon ◽  
Salmonella Typhi ◽  
Antibacterial Resistance ◽  
Human Colon Cancer ◽  
Antibacterial Drugs ◽  
Excoecaria Agallocha ◽  
Extract Fraction

Abstract Background The incidence of antibacterial resistance and colon cancer is increasing in India. Antibacterial resistance and chemoresistance demand the need of developing herbal or natural chemotherapeutic agents. Our study thus, aims to determine the antibacterial and anticancer activities of the leaf extracts of the mangrove Excoecaria agallocha. Results Liquid chromatography–mass spectroscopy analysis of the ethanolic E. agallocha extracts revealed the presence of Bergenin. The plant extract fraction containing Bergenin had potent antibacterial action against a resistant strain of Salmonella typhi with an MIC value of 15.7 ± 0.04 µg/mL. Treatment of the bacteria with the plant extract made it moderately susceptible to the antibacterial drugs ampicillin, aztreonam, cefotaxime, chloramphenicol and imipenem. The plant extract caused membrane damage and disrupted the expression of a 33 kDa outer membrane protein (OmpA) in S. typhi. It was plausibly due to this mechanism of the plant extract that made the bacteria susceptible to the antibacterial drugs to a certain extent. Further, fluorescence microscopy analysis revealed the anticancer property of the extract against a human colon cancer (DLD-1) cell line by activation of Caspase-3followed by subsequent apoptosis and exhibited cytotoxicity against the cancerous cell line with an IC50 value of 17.99 ± 1.12 µg/mL. Caspase-3 activity was observed to increase in a dose-dependent manner as determined by spectrophotometric assays. Moreover, the expression of the metalloproteinase-7 (MMP-7) was significantly reduced in plant extract treated DLD-1 colon cancer cells. Conclusion The results indicate that E. agallocha is a novel source of Bergenin, and the plant extract fraction under study may be used in combination therapy along with antibacterial drugs to combat antibacterial resistance of S. typhi and also to alleviate the risks of colon cancers in human. However, further investigations may be undertaken for its therapeutic application and to explore its potential bioactivity against other bacterial strains and human cancer cell lines.

scholarly journals Polypeptide Fraction fromArca subcrenataInduces Apoptosis and G2/M Phase Arrest in HeLa Cells via ROS-Mediated MAPKs Pathways

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
Xianjing Hu ◽  
Zhang Zhang ◽  
Ting Liu ◽  
Liyan Song ◽  
Jianhua Zhu ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Hela Cells ◽  
Underlying Mechanism ◽  
Cyclin B1 ◽  
Mitochondrial Pathway ◽  
Dependent Manner ◽  
Phase Arrest ◽  
M Phase ◽  
Induced Apoptosis ◽  
High Level

Arca subcrenatais documented in the literature of marine Traditional Chinese Medicine. Polypeptide fraction fromA. subcrenata, coded as P2, was demonstrated to possess significant antitumor activity in our previous study. However, the underlying mechanism remains undefined. The present study was carried out to investigate the underlying antitumor mechanism of P2 in human cervical cancer HeLa cells by MTT, FCM, LSCM, and western blot assays. The results revealed that P2 significantly induced apoptosis of HeLa cells in a concentration- and time-dependent manner. High level of ROS was provoked by P2, which was in turn responsible for induction of apoptosis through activation of intrinsic mitochondrial pathway and JNK1/2, p38 MAPK pathways, as well as inhibition of ERK1/2 pathway, as evidenced by the abrogation of P2’s effect on HeLa cells preincubated with the ROS scavenger NAC. P2 also was observed to display significant effect on G2/M phase arrest by downregulating the expression of cyclin B1/cdc2 complex and upregulating the expression of p21. These findings demonstrate that P2 induces apoptosis and G2/M phase arrest in HeLa cells through ROS-mediated MAPKs pathways, suggesting that P2 would be worth investigating as a promising agent within the scope of marine drugs for treatment of cervical cancer.

scholarly journals ZNF582 Methylation Predicts Better Radiotherapy Response of Cervical Cancer and ZNF582 Overexpression Reduces Radiosensitivity by Arresting the Cell Cycle in S Phase

2021 ◽  
Author(s):  
Nayiyuan Wu ◽  
Xiaoyun Zhang ◽  
Miaochen Zhu ◽  
Chao Fang ◽  
Xiaoting Liu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Cell Cycle Arrest ◽  
Locally Advanced ◽  
Underlying Mechanism ◽  
S Phase ◽  
Cycle Arrest ◽  
Cervical Cancer Cells ◽  
Pre Treatment ◽  
The Relationship

Abstract Purpose: Our previous studies identified ZNF582 methylation (ZNF582m) level as a useful biomarker for cervical cancer screening, detection, and prognosis. The purpose of this study is to investigate the relationship between ZNF582m level and cervical cancer radiotherapy sensitivity and its underlying mechanism.Patients and Methods: This was a prospective multicenter clinical study, included two independent cohorts locally advanced cervical cancer patients. Exfoliated cervical cancer cells were collected pre-treatment and during treatment (after 24, 30, 36, 48, and 64 Gy) to test ZNF582m level, radiotherapy response evaluated according to RECIST Version. Results: In the first cohort, 22 cases achieved satisfied response, 28 exhibited modest response. Radiotherapy reduced ZNF582m level among all patients. Baseline ZNF582m was significantly higher in the satisfied response cases than in modest response cases, also, patients with high baseline ZNF582m (ZNF582m-high, n = 21) were more sensitive to radiotherapy than ZNF582m-low patients (n = 29), as evidenced by greater satisfied response rate (76.2% vs. 20.7%). An independent cohort confirmed above results. The magnitude of ZNF582m reduction was associated with a radiotherapeutic response, a subset of ZNF582m-low patients (5 of 28) exhibiting a transient increase in ZNF582m demonstrated greater radiosensitivity than other ZNF582m-low patients. ZNF582 overexpression induced cell cycle arrest in S phase.Conclusion: High ZNF582m level predicts better cervical cancer radiosensitivity, ZNF582 overexpression reduces radiosensitivity by cell cycle arrest.

Comparison of traditional Chinese medicine (TCM) herbal mixture LQ and paclitaxel on the phase of cancer cell cycle blockage and cancer cell invasion.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e22014-e22014
Author(s):  
Robert M. Hoffman ◽  
Chengyu Wu ◽  
Shuya Yano ◽  
Lei Zhang
Keyword(s):  
Cell Cycle ◽  
Cancer Cell ◽  
Hela Cells ◽  
Positive Control ◽  
Control Group ◽  
Herbal Mixture ◽  
Cycle Arrest ◽  
Anti Cancer ◽  
M Phase ◽  
Treated Culture

e22014 Background: Although Traditional Chinese Medicine (TCM) has been used to treat cancer for thousands of years, the mechanisms of action are either poorly understood or unknown. In the present report, we use state-of-the-art technology to investigate the anti-cancer mechanism of the TCM herbal mixture LQ. Methods: Fluorescence ubiquitination-based cell cycle indicator (FUCCI) was used to monitor cell cycle arrest HeLa cells after LQ treatment. FUCCI-HeLa cells were cultured in two dimensional (2D) monolayer, Matrigel and 3D Gelfoam. Changes of cell cycle status were observed using the Olympus FV1000 confocal imaging system whereby cycling cells fluoresce green and quiescent cells red. Paclitaxel (Taxol) was used as the positive control. Results: Paclitaxel induced a G2/M cell cycle block. In contrast, LQ blocked FUCCI-HeLa cells in the G0/G1 phase of the cell cycle in all 3 culture models. In monolayer culture, the paclitaxel positive control had approximately 45% of the cells in G2/M phase. In contrast, the LQ-treated cells were mostly in the G0/G1 phase (>90%). In Matrigel culture, HeLa cells formed spheres. The spheres in the paclitaxel control group had 40% of the cells in G2/M phase, but only 15% in LQ-treated cultures. In 3D Gelfoam culture, cells grew along the structures of the Gelform. The paclitaxel positive control culture had approximately 45% of cells in G2/M phase. In contrast, the cells in LQ-treated culture were mostly in G0/G1 phase (>80%). The cells in pacilitaxel control group invaded to 250~300 µm deep in the Gelfoam, but only 150~200 µm deep in LQ-treated culture. Conclusions: The anti-cancer mechanism of TCM herbal mixture LQ involves cancer cell cycle arrest at G0/G1 and inhibition of cancer cell invasion.

scholarly journals Role of NRF2 cascade in determining the differential response of cervical cancer cells to anticancer drugs- an in vitro study

2021 ◽  
Author(s):  
Pushkal Sinduvadi Ramesh ◽  
Sharanya Raja ◽  
Shwethambari Harave Udayakumar ◽  
Shruthi Chandrashekar ◽  
Suma M Nataraj ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Enzyme Activity ◽  
Cancer Cells ◽  
Anticancer Drugs ◽  
Differential Response ◽  
Cancer Drugs ◽  
Nrf2 Pathway ◽  
Siha Cells ◽  
Anti Cancer ◽  
Cervical Cancer Cells

Abstract Background Cervical cancers are usually treatable if detected in early stages by a combination of therapies. However, the prognosis of cervical cancer patients with metastasis remains unfavorable due to the fact that most of the cervical carcinomas are either resistant to anticancer drugs or show signs of relapse after initial treatment. Therefore, it is important to control the chemoresistance as it is the key to develop effective treatment options for cervical cancer. Objective The current study aimed at evaluating the differential responses of cervical cancer cells to anti-cancer drugs and assessed whether the differences in the expression profiles of antioxidant genes regulated by NRF2 (nuclear factor erythroid-2-related factor 2), led to the variations in the sensitivities of the cancer cells to treatment. Methodology: Three cervical cancer cell lines were investigated for their differences in NRF2 pathway by measuring the gene expression and enzyme activity. The differences in the sensitivity to anti-cancer drugs and variation in ROS profile was also evaluated. The addition of exogenous drugs to manipulate the intracellular ROS and its effect on NRF2 pathway genes was also investigated. Results HeLa and SiHa cells were more sensitive to cisplatin and oxaliplatin treatment than C33A cells. HeLa and SiHa cells had significantly lower NRF2 gene levels, NQO1 enzyme activity and basal GSH levels than C33A cells. Levels of ROS induced were higher in HeLa than C33A cells. Conclusion Overall, the differences in the cellular levels of antioxidant regulatory genes led to the differential response of cervical cancer cells to anti-cancer drugs.

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