Synergistic effect of sorafenib with Platycladus orientalis (L) leaf extract on cervical cancer

The extracts of Platycladus orientalis (L.) Franco leaves have shown promising anti-cancer, anti-oxidant and anti-inflammatory potency with the traditional knowledge of healing HPV associated warts. The purpose of this research is to assess the synergistic activity of sorafenib and Platycladus orientalis (L) leaf extraction on cervical cancer cells. The cytotoxicity efficiency of different concentrations of Sorafenib and ethanol extract of Platycladus orientalis (L.) leaves were tested on HeLa cells by MTT and Trypan blue assays. The synergistic effect of the IC50 concentrations of Sorafenib and Platycladus orientalis (L.) on HeLa cell by MTT assay, and mRNA expression levels of tumor suppressor tazarotene-induced gene 3 (TIG3), proliferating cell nuclear antigen (PCNA) gene and apoptosis modulator (Bcl-2) gene by RT-PCR were evaluated with individual treatments. Combination treatment showed a relatively more expression of TIG3 and less expression of Bcl-2 and PCNA was observed. Growth factor-induced MAPKP activation was arrested by compound combination treatment, which and suppression of proliferation-induced apoptosis of cervical cancer cells. Based on the our results, the combination of sorafenib and crude leaf extract from Platycladus orientalis (L.) can effectively suppress cervical cancer cell growth, thereby providing an interesting rationale for further clinical trials and in-vivo studies.

Selective photodynamic effects on cervical adenocarcinoma cells provided by F127 Pluronic®-based micelles modulating hypericin delivery

Abstract Cervical cancer is a leading cause of death among women. The endocervical adenocarcinoma (ECA) represents an aggressive and metastatic type of cancer with no effective treatment options currently available. We evaluated the antitumoral and anti-migratory effects of hypericin (HYP) encapsulated on Pluronic F127 (F127/HYP) photodynamic therapy (PDT) against a human cell line derived from invasive cervical adenocarcinoma (HeLa) compared to a human epithelial cell line (HaCaT). The phototoxicity and cytotoxicity of F127/HYP were evaluated by the following assays: colorimetric assay, MTT, cellular morphological changes by microscopy and long-term cytotoxicity by clonogenic assay. In addition, we performed fluorescence microscopy to analyze cell uptake and subcellular distribution of F127/HYP, cell death pathway and reactive oxygen species (ROS) production. The PDT mechanism was determined with sodium azide and D-mannitol and cell migration by wound-healing assay. The treatment with F127/HYP promoted a phototoxic result in the HeLa cells in a dose-dependent and selective form. Internalization of F127/HYP was observed mainly in the mitochondria, causing cell death by necrosis and ROS production especially by the type II PDT mechanism. Furthermore, F127/HYP reduced the long-term proliferation and migration capacity of HeLa cells. Overall, our results indicate a potentially application of F127/HYP micelles as a novel approach for PDT with HYP delivery to more specifically treat ECA.

CRISPR/Cas9-mediated Knockout of KIFC1 Inhibits Proliferation and Induces Apoptosis of Cervical Cancer Cells / 肿瘤防治研究

Objective To investigate the functions of the KIFC1 gene in tumor cells and its effect on the proliferation of cervical cancer cells. Methods We designed sgRNAs targeting the KIFC1 gene and constructed a recombinant plasmid based on the pSpCas9 (BB)-2A-GFP vector, which was co-transfected into HeLa cells. We screened monoclonal knockout cell lines through flow cytometry sorting, limited dilution inoculation of cells, and sequencing. RT-qPCR, Western blot, and immunofluorescence were used to detect the transcription and protein expression levels of KIFC1 in knockout cells. Cell phenotypes such as nucleus and microtubule cytoskeleton were observed using phase-contrast microscopy and fluorescence confocal microscopy. Cell proliferation, cell cycle, and apoptosis were analyzed by growth curve plotting, EdU labeling, and acridine orange staining. Results The deletion of the KIFC1 gene resulted in the abnormal phenotypes of HeLa cells, with increased numbers of multinuclei, micronucleus, and disordered microtubules. The cell cycle was disrupted, accompanied with a significant increase in the ratio of late apoptotic cells and a decrease in cell proliferation (all P < 0.05). Conclusion KIFC1 gene deletion affects the assembly of microtubules and cell division in HeLa cells, leading to abnormal nuclear morphology, chromatin elimination, cell cycle arrest, and increased cell apoptosis.

Arracacia xanthorrhiza Bancr compounds decrease β-actin, hypoxia-inducible factor-1 and nitric oxide production in HeLa cells / Los componentes de la Arracacia xanthorrhiza Bancr disminuyen las producciones de la actina β, del factor-1 inducible por la hipoxia y del óxido nítrico en células Hela

Abstract The treatment of cancer patients with anti-cancer drugs is often accompanied by the presence of undesirable side effects. The use of natural plant derivatives alone, or in conjunction with existing anti-neoplastic drugs, has been suggested to obtain better results and decrease these side effects. Nitric oxide (NO•), the hypoxia-inducible factor-1 (HIF-1), and decreased concentration of actin play important roles in cancer progression. The beneficial effects of polyphenols in various organ disorders including cancer has been reported. The aim of this study was to determine the effect of Arracacia xanthorrhiza Bancr extracts, white (WAXB) and red (RAXB) variants (compounds rich in polyphenols) on the concentrations of β-actin, NO• and HIF-1 in Hela cells cultures, to uncover possible anti-neoplastic effects. Extracts from the plant leaves were added to Hela cell cultures at a concentration of 10-3 mg/mL, and after 24 hours of culture, the concentrations of β-actin, NO• and HIF-1 were determined by immunohistochemical, biochemical and western blot assays. Both extracts reduced the concentrations of β-actin, NO• and HIF-1 (p<0.001), similar to the methotrexate effect. These results suggest an antineoplastic effect of the studied plant extracts and highlight the possibility of their use in the treatment of neoplasms.

Resumen El tratamiento de pacientes con cáncer utilizando drogas-antineoplásicas presenta problemas relacionados con efectos colaterales indeseables. Se ha sugerido el uso de derivados de plantas naturales solas, o en combinación con drogas antineoplásicas existentes para obtener mejores resultados y disminuir los efectos colaterales. Así mismo, se ha reportado que el óxido nítrico (NO•), el factor-1 inducible por hipoxia (HIF-1) y la disminución de la expresión de la actina tienen un papel en la progresión del cáncer. También se ha reportado los efectos beneficiosos de lo polifenoles en varios desordenes orgánicos, incluyendo el cáncer. El objetivo de este estudio fue determinar los efectos de los extractos procedentes de la Arracacia xanthorrhiza Bancr blanca (AXBB) y la variedad roja (AXBR) (compuestos ricos en polifenoles) en las concentraciones de la actina-beta, el NO• y el HIF-1 en cultivo de células Hela, para destacar sus posibles efectos antineoplásicos. A los cultivos de células Hela se les agregaron los extractos de las hojas de AXBB o AXBR (10-3 mg/mL, concentración final) y después de 24 horas de cultivo se determinaron las concentraciones de la actina-beta, el NO• y el HIF-1 por métodos inmunohistoquímicos, bioquímicos y western blot. Ambos extractos disminuyeron las concentraciones de la actina-beta, el NO• y el HIF-1 (p<0,001) de una manera similar al efecto del metotrexato. Estos resultados sugieren un efecto antineoplásico de estos extractos y destacan la posibilidad de ser usados para el tratamiento de las neoplasias.

Effects of SP600125 on Proliferation and Invasion of Human Cervical Cancer HeLa Cells / 肿瘤防治研究

Objective To investigate the effect of SP600125 on the proliferation, cell cycle, apoptosis and invasion of human cervical cancer HeLa cells. Methods CCK-8 method was used to detect the proliferation of HeLa cells treated with different concentrations of SP600125 at different time points. The 20 μmol/L of SP600125 was determined for subsequent experiments. Cell proliferation ability was detected using plate clone formation assay; nuclear morphology was observed by DAPI staining; cell cycle and apoptosis were measured by flow cytometry; cell migration and invasion were detected by cell scratch and Transwell methods; the mRNA and protein levels of p53, Mad2L1 and CDC20 were measured by qRT-PCR and Western blot after SP600125 treatment at different time points. Results Compared with control group (0.1%DMSO), cells proliferative activity were reduced by 10, 20, 30, 40 and 50 μmol/L SP600125 treatment for 24h. Compared with control group, the rate of apoptosis was significantly increased in SP600125 treatment groups, and the cell proportion in G2/M phase increased (P < 0.001), while the cell proportion in G0/G1 phases cells was reduced after SP600125 treatment for 24h and 48h (P < 0.001), and the clonal number, migration and invasion ability of HeLa cells also decreased significantly (P < 0.001). qRT-PCR and Western blot results showed a significant decrease in Mad2L1 mRNA and protein expression (P < 0.05) and a significant increase in p53 and CDC20 mRNA and protein expression (P < 0.01). Conclusion SP600125 can induce cell cycle arrest of cervical cancer HeLa cells in G2/M phase by upregulating p53 and CDC20 and downregulating Mad2L1 expression, and promote cell apoptosis to inhibit cell proliferation, migration and invasion.

Juglone induces proliferation inhibition and apoptosis of cervical cancer cells via promoting c-Myc ubiquitination / 南方医科大学学报

OBJECTIVE@#To observe the expression of c-Myc protein in cervical cancer HeLa cells and explore the effect of juglone on the proliferation and apoptosis of HeLa cells by affecting c-Myc ubiquitination.@*METHODS@#HeLa cells treated with different concentrations (0, 10, 20, or 50 μmol/L) of juglone or with 20 μmol/L juglone for different time lengths were examined for expression of c-Myc protein with Western blotting. The half-life of c-Myc protein was determined using cycloheximide (CHX) and c-Myc protein degradation was detected using coimmunoprecipitation. We also assessed the effects of 20 μmol/L juglone combined with 0, 1.0 or 2.0 μmol/L MG132 (a proteasome inhibitor) on c-Myc expression. The effects of 20 μmol/L juglone on the proliferation and apoptosis of HeLa cells with RNA interference-mediated knockdown of c-Myc were evaluated with MTT assay and flow cytometry.@*RESULTS@#Treatment with juglone significantly lowered c-Myc protein expression in HeLa cells in a concentration-and time-dependent manner (P < 0.05). Juglone obviously shortened the half-life of c-Myc protein, and the addition of MG132 significantly up-regulated the expression level of c-Myc protein (P < 0.05). Juglone treatment also promoted ubiquitination of c-Myc protein in HeLa cells. Compared with the cells transfected with a negative control construct, the cells transfected with si-c-Myc showed significantly decreased proliferation inhibition and a lowered cell rate with early apoptosis after juglone treatment (P < 0.05).@*CONCLUSION@#Juglone inhibits proliferation and promotes apoptosis of HeLa cells by affecting the ubiquitination of c-Myc protein.

Enhanced Effect of miR-148a Targeting STAT3 on Chemosensitivity of Cervical Cancer HeLa Cells to Cisplatin / 肿瘤防治研究

Objective To investigate the effect of miR-148a on the chemosensitivity of cervical cancer HeLa cells to cisplatin and its related mechanism. Methods Cervical cancer HeLa cells were cultured in vitro and the concentration gradient of cisplatin was set to detect IC20 value. Control group, mimic control group, miR-148a mimic group, inhibitor control group and miR-148a inhibitor group were set up. qRT-PCR was used to detect the expression of miR-148a and STAT3 mRNA after transfection. After 4 μmol/L cisplatin treatment, the proliferation of HeLa cells was detected by MTT assay; the apoptosis and cell cycle distribution were detected by flow cytometry; Western blot was used to detect the protein expression of p-STAT3/STAT3, CyclinD1, Bcl-2, Bax and Cleaved caspase-3. Results The IC20 of cisplatin on HeLa cells was about 4 μmol/L. Compared with the mimic control group, the level of miR-148a in the miR-148a mimic group was significantly increased, and the level of STAT3 mRNA was significantly decreased (P < 0.05). Compared with the inhibitor control group, the level of miR-148a in HeLa cells in miR-148a inhibitor group was significantly decreased, and the level of STAT3 mRNA was significantly increased (P < 0.05). On the basis of cisplatin treatment, compared with the mimic control group, the apoptosis rate, G0/G1 phase cell ratio, the protein levels of Bax and Cleaved caspase-3 were significantly increased in miR-148a mimic group, while OD value, the proportions of cells in S and G2/M phase, the protein levels of p-STAT3/STAT3, CyclinD1, Bcl-2 were significantly decreased (P < 0.05); compared with the inhibitor control group, the above indicators of HeLa cells in miR-148a inhibitor group changed significantly in the opposite direction (P < 0.05). Conclusion MiR-148a could enhance the chemosensitivity of cervical cancer HeLa cells to cisplatin by targetedly inhibiting STAT3.

Combination Effect of Tamoxifen and Ascorbic Acid Treatment on Breast Cancer Cells (MCF-7) and Cervical Cancer Cells (HeLa) Kesan Rawatan Kombinasi Tamoksifen dan Asid Askorbik ke atas Sel Kanser Payudara (MCF-7) dan Sel Kanser Serviks / Jurnal Sains Kesihatan Malaysia

@#Breast cancer and cervical cancer are among the leading causes of death among women in the world. Even though chemotherapy is available as cancer treatment, the development of drug resistance in both cancer cells has reduced the efficacy of chemotherapeutic drugs in such treatment. The current study was aimed to evaluate the cell viability of human breast cancer cells, MCF-7, and cervical cancer cells, HeLa upon the combination treatment of ascorbic acid and tamoxifen. The cell viability was measured using the MTT assay, with an incubation period of 72 hours in a humidified CO2 incubator. The concentrations of tamoxifen and ascorbic acid that reduced 50% of the cell population (IC50) were determined from the dose-response curve. The IC50 concentration was used to determine the cell viability in the treated cells. CompuSyn software was used to evaluate the combined effects towards both cells upon treatment and the results were calculated as combination index (CI). The data were analyzed using GraphPad Prism (version 7). Statistical analysis was performed using an independent t-test. The IC50 values of tamoxifen and ascorbic acid on MCF-7 cells were 14.53 µg/ml and 15.8 µg/ml respectively, while the IC50 values of tamoxifen and ascorbic acid on HeLa cells were 11.09 µg/ml and 202.3 µg/ml respectively. The combination of tamoxifen and ascorbic acid exerted a greater growth reduction percentage in both cells compared to tamoxifen alone. The results indicated that ascorbic acid synergizes the cytotoxic effect of tamoxifen at lower concentrations towards MCF-7 cells with a CI less than 1. However, the combination of tamoxifen and ascorbic acid exerted an antagonistic effect in HeLa cells, with a CI more than 1.

Preparation of paclitaxel-loaded and folic acid-modified poly (lactic-co-glycolic acid) nano-micelles and in vitro anticancer effect on cervical cancer HeLa cells / 中国中药杂志

The paclitaxel-loaded and folic acid-modified poly(lactic-co-glycolic acid) nano-micelles(PTX@FA-PLGA-NMs) were prepared by the emulsion solvent evaporation method, and the parameters of paclitaxel-loaded nano-micelles were optimized with the particle size and PDI as evaluation indexes. The morphology of the nano-micelles was observed by transmission electron microscopy(TEM), and the stability, drug loading and encapsulation efficiency were systematically investigated. In vitro experiments were performed to study the cytotoxic effects of nano-micelles, apoptosis, and cellular uptake. Under the optimal parameters, the nano-micelles showed the particle size of(125.3±1.2) nm, the PDI of 0.086±0.026, the zeta potential of(-20.0±3.8) mV, the drug loading of 7.2%±0.75%, and the encapsulation efficiency of 50.7%±1.0%. The nano-micelles were in regular spherical shape as observed by TEM. The blank FA-PLGA-NMs exhibited almost no inhibitory effect on the proliferation and growth of tumor cells, while the drug-loaded nano-micelles and free PTX exhibited significant inhibitory effects. The IC_(50) of PTX@FA-PLGA-NMs and PTX was 0.56 μg·mL~(-1) and 0.66 μg·mL~(-1), respectively. The paclitaxel-loaded nano-micelles were potent in inhibiting cell migration as assessed by the scratch assay. PTX@FA-PLGA-NMs had good pro-apoptotic effect on cervical cancer HeLa cells and significantly promoted the uptake of HeLa cells. The results of in vitro experiments suggested that PTX@FA-PLGA-NMs could target and treat cervical cancer HeLa cells. Therefore, as nanodrug carriers, PTX@FA-PLGA-NMs with anti-cancer activity are a promising nano-system for improving the-rapeutic effects on tumors.

Study on Reversal Effects of Levoshikonin on Cisplatin Resistance of Human Cervical Carcinoma HeLa Cells / 中国药房

OBJECTIVE：To investigate the reversal effects and potential mechanism of levoshikonin （L-SHK）on cisplatin （DDP）resistance of human cervical carcinoma HeLa cells. METHODS ：Human cervical carcinoma HeLa cells were used as research objects ，and drug-resistant HeLa/DDP cells were induced by DDP. CCK- 8 assay was used to determine drug resistance index of HeLa/DDP cells ，the inhibition rate of different doses of L-SHK （0.125，0.25，0.5，1，2，4，8，16 μmol/L）on cell proliferation，IC50 and the reversion index of L-SHK on HeLa/DDP cells. Effects of low ，medium and high doses of L-SHK （0.3， 0.6，1.2 μmol/L）combined with DDP on cell cycle and apoptosis rate were determined by flow cytometry. Western blotting assay was used to detect the effects of low ，medium and high doses of L-SHK （0.3，0.6，1.2 μmol/L）combined with DDP on the expression of apoptosis-related protein （Cleaved caspase- 3，Bcl-2 and Bax ）. RESULTS ：The drug resistance index of HeLa/DDP cells was 11.8. The inhibition rate of L-SHK on HeLa/DDP cells increased with the increase of dose. Compared with DDP alone ， IC50 of DDP+low-dose ，medium-dose and high-dose L-SHK groups were decreased significantly ，with a dose-dependent manner （P＜0.05）. The reversion indexes were 1.38，2.80，6.71 in DDP+low-dose ，medium-dose and high-dose L-SHK groups. Compared with blank control group ，the proportion of cells at phase G 0/G1 and phase S in administration groups ，as well as early and late apoptosis rate and total apoptosis rate of cells ，the protein expression of Bax and Cleaved caspase- 3 in L-SHK combination groups were increased significantly ；the proportion of cells at phase G 2/M in administration group as well as the protein expression of Bcl-2 in L-SHK combination groups were decreased significantly （P＜0.05）. Compared with DDP group ，the proportion of cells at phase S and G 2/M and the protein expression of Bcl- 2 in L-SHK combination groups were significantly decreased ；the proportion of cells at phase G 0/G1，early and late apoptosis rate and total apoptosis rate ，the protein expression of Bax and Cleaved caspase- 3 in L-SHK combination groups were significantly increased （P＜0.05）. CONCLUSIONS ：HeLa/DDP cells are resistant to DDP ，and L-SHK can reverse the drug resistance. L-SHK combined with DDP can promote the apoptosis of HeLa/DDP cells ，which is better than DDP alone. Its mechanism may be related to the influence of cell cycle and the regulation of apoptosis-related protein expression.

Effect of Paiteling on HeLa Cell Proliferation and Metastasis Ability and PI3K/Akt Signal Transduction Pathway / 中国实验方剂学杂志

Objective:To explore the effect of Paiteling on the proliferation，metastasis and invasion of HeLa cells and relevant proteins of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway. Method:① HeLa cells were divided into blank group and Paiteling concentration gradient groups (3.906，2.604，1.953，1.563，1.302，1.116，0.977 g·L-1). After drug intervention for 24 h，the cell morphological changes were observed under microscope. The cell viability was measured by thiazole blue (MTT) colorimetry，and the half inhibitory concentration (IC50) of Paiteling on HeLa cells was calculated. ② HeLa cells were divided into blank group，cisplatin group (0.01 g·L-1)，Paiteling high-dose group (2.974 g·L-1)，Paiteling medium-dose group (1.487 g·L-1) and Paiteling low-dose group (0.991 g·L-1). Cell proliferation and toxicity test (CCK-8) method was used to detect the effect of Paiteling on the proliferation ability of HeLa cells，scratch test was used to detect cell migration，and invasion test (Transwell) was used to detect changes in cell invasion ability. ③ Inhibitor LY294002 group (0.006 g·L-1) was added. Western blot (WB) was used to detect the expressions of Paiteling on PI3K，Akt，recombinant human B-cell lymphoma factor-xl (Bcl-xl)，and B-cell lymphoma/leukemia associated D protein (Bad). Result:① Compared with the blank group，microscopic observation showed that the number of cells in the treatment group was significantly reduced, and the cell morphology was incomplete. MTT experiments showed that Paiteling has a significantly inhibitory effect on HeLa cell proliferation (P<0.01). The IC50 of Paiteling on HeLa cells was calculated as 2.974 g·L-1. ② The CCK-8 experiment showed that compared with the blank group，all the drug-treated groups had an inhibitory effect on HeLa cell proliferation at 24，36，48 h (P<0.01), compared with the cisplatin group，middle and low-dose Paiteling groups showed a reduced inhibitory effect on HeLa cell proliferation at each time point (P<0.01). The scratch test showed that，compared with the blank group，each drug-added group could inhibit the migration ability of HeLa cells (P<0.01)，and the cell migration rate of the high-dose Paiteling group was lower than that of the cisplatin group (P<0.05). Transwell experiments showed that compared with the blank group，the number of membranes permeated by HeLa cells in each drug-treated group was decreased (P<0.01)，and the number of membranes permeated in the middle and low-dose Paiteling groups was increased compared with the cisplatin group (P<0.01). ③ Western blot showed that compared with the blank group，the expression levels of PI3K，Bcl-xl，and Akt in the high，medium，and low-dose Paiteling groups and the LY294002 group decreased (P<0.05，P<0.01)，while the expression of Bad increased (P<0.01). Compared with the high-dose Paiteling group，the PI3K，Akt，and Bcl-xl protein expressions were increased in the low-dose Paiteling group (P<0.01)，whereas Bad expression was decreased (P<0.01). Conclusion:Paiteling can inhibit HeLa cell proliferation，metastasis and invasion ability in a dose-dependent and time-dependent manner，which may be related to its effect on the expressions of PI3K/Akt signaling pathway-related proteins.

Fermentation conditions and antitumor products of an ocean-derived fungus Alternaria sp. 114-1G: a preliminary investigation / 国际药学研究杂志

Objective: To investigate and optimize the fermentation conditions of an ocean-derived fungus, Alternaria sp. 114- 1G, and isolate metabolites in the fermentation products to explore antitumor compounds in the products via the structure elucidation and in vitro antitumor activity assay. Another aim of this study is to accomplish a fundamental work for further research on new compounds from Alternaria sp. 114- 1G. Methods: The in vitro antitumor activity was assayed for the crude extracts and isolated compounds by the CCK-8 method using a human cervical cancer HeLa cell line. The fermentation conditions were investigated and opti- mized based on the biomass of the fermentation and the in vitro antitumor activity of crude extracts of the fermentation. For the separation and isolation of metabolites, the column chromatography was performed on silica gel and Sephadex LH-20 columns, and semi-preparative high performance liquid chromatography(semi-HPLC)was conducted on a COSMOSIL C18-MS-Ⅱ column(10 mm×250 mm). The obtained compounds were identified according to the MS, 1H NMR and13C NMR data. Results: The relatively favored fermentation conditions were as follows: mannitol 25 g/L, maltose 15 g/L, glucose 10 g/L, monosodium glutamate 10 g/L, soybean peptone 5 g/L, yeast extract 3 g/L; 60% sea water in whole medium, initial pH 7.5; and fermentation at 10℃ for 15 days under a 150 r/min shaking speed on a rotary shaker. Four compounds 1-4 were isolated from the fermentation products of 114-1G strain and identified as cyclo (Gla-Tyr)(1), cyclo(Ala-Ile)(2), thymidine DNA nucleotide(3)and pachybasin(4). Among them, 4 showed a relatively stronger inhibitory activity on HeLa cells, with the 57.8% of inhibition rate at 100 μg/ml. Some other compounds were also isolated, and a phenolic one had been shown to be a new compound by a literature survey according to the planar structure deduced. Further studies on their structures were in progress. Conclusion: Fermentation conditions for Alternaira sp. 114-1G were investigated preliminarily, and four compounds 1-4 have been isolated from the fermentation products of the 114-1G strain. Among them, pachybasin(4)showed a relatively higher inhibitory effect on human cancer HeLa cells. Alternaira sp. 114-1G could produce new metabolites and the present study has provided a reliable groundwork for further research on new compounds from Alternaira sp. 114-1G.

Identification And Cytotoxic Activity From N– Hexane Fraction Of Bruguiera Gymnorhiza To Hela Cells Against

introduction: Bystander Cardiopulmonary Resuscitation (CPR) is a vital life-saving intervention, easy skills to learn and performed when witnessing a victim of cardiopulmonary arrest. since the School teachers are the rst line of interventions in schools where students spend 7 to 8 hours per day thus any life-threatening situation could happen so, School teachers must be aware of BLS performance. this study conducted to assess the awareness and attitude of School teachers regarding BLS. Methodology An analytic cross-sectional study through self-administered modied questionnaire among 469 from school teachers in Qatif, Dammam and Khobar city, Eastern Province, Saudi Arabia. the questionnaire was validated by 8 experts from family medicine experts, public health and department of emergency, The questionnaire is divided into ve main parts as follows: demographic data section ,Knowledge towards BLS performance ability, Attitude towards BLS. And Barriers. Results: only 23.8 % of school teachers had previous training, about half of school teachers had poor BLS knowledge. 25.8% Knew the contact numbers for emergency ,44.4% knew what the rst step in BLS. only 14.1% are competent to do BLS if indicated. Majority of secondary school teachers are willing to enroll in BLS courses (92.9%) and Majority of them (90.7%) agree that BLS should be a part of school curriculum . in another hand only (35.3%) agreed that BLS training should be mandatory to their job's certication. conclusion:Although a majority of teachers not received previous CPR training, and had poor BLS knowledge, willingness to have BLS training was high and having BLS as a part school's curriculum was recommended by the majority of the teachers

Cytotoxicity Activity and Phytochemical Screening of Anthocleista djalonensis Root Extracts against Cancer Cells

Aim: Several medicinal uses have been reported for Anthocleista djalonensis and many types of pure compounds have been isolated. However, the anti-cancer activity of this plant has not been proven. The aim of this study was to screen for the phytochemicals present in the rootn-hexane, ethyl acetate, and acetone extracts of Anthocleista djalonensis, and to evaluate its anticancer potential against human cervix adenocarcinoma cells (HeLa cells) in vitro. Place and Duration of Study: The study was carried out in Department of Organic Chemistry, Rhodes University, Grahamstown, South Africa. The duration period was between March and July, 2016. Methodology: Extracts were prepared by soaking the root powder in the respective solvents with continuous stirring; The extracts were filtered and evaporated to remove the solvents. The extracts were then screened for phytocompounds by preliminary screening methods. Anti-cancer potential was carried out by a Resazurin assay and CC50 values were determined. Results: The extracts showed the presence of carbohydrates, glucoside, alkaloids, flavonoids, terpenoids, tannins, saponins, sterols. All extracts demonstrated moderate cytotoxicity against HeLa cells. Conclusion: The hexane, ethyl acetate and acetone extracts showed anticancer property. The roots extracts of Anthocleista djalonesis were thus found to possess potential anticancer activities.

Measles virus phosphoprotein inhibits apoptosis and enhances clonogenic and migratory properties in HeLa cells

Measles virus is the causative agent of measles, a major cause of child mortality in developing countries. Two majorproteins, coded by the viral genome, are nucleocapsid protein (N) and phosphoprotein (P). The N protein protects the viralgenomic RNA and forms ribonucleoprotein complex (RNP) together with P protein. MeV-P protein recruits the largeprotein (L), i.e. viral RNA-depended RNA polymerase (RdRp), to ensure viral replication in host cell. Apoptogenicproperties of N protein of Edmonston vaccine strain have been established in our lab previously. We investigated the role ofMeV-P protein of Edmonston vaccine strain as modulator of apoptosis in cervical cancer cell line (HeLa) and found thatMeV-P protein is anti-apoptotic and enhances cell proliferation. Measles virus is considered to be innately oncotropic virus.However, the anti-apoptotic property of MeV-P protein raises important concerns while adopting this virus as an anti-cancertherapeutic tool.

The extraordinary trajectory of the Papillomavirus through human history / A extraordinária trajetória do Papilomavirus através da história da humanidade

Papillomaviruses have been infecting people since the beginning of human life on earth. The most relevant chapters of this story were written by the brilliant, diverse, and prominent scientists of their respective times. However, an important part of this story is the Papillomavirus victims and their adverse situations. There have also been disputes over intellectual primacy of the discoveries and the collaborators who were not given the recognition according to the role they played. This article will guide the reader through the remarkable facts of this conflicting and interesting relationship between humans and the Papillomavirus

O papilomavírus tem infectado pessoas desde o começo da vida humana na Terra. Os capítulos mais relevantes dessa história foram escritos por mãos brilhantes de diversos e proeminentes cientistas em seus respectivos tempos. No entanto, parte importante dessa história também foi construída por vítimas do papilomavírus e suas situações adversas. Houve também disputas sobre a primazia intelectual das descobertas e os colaboradores cujo reconhecimento é menor que o papel que desempenhavam. Este artigo levará o leitor por um breve passeio pelos fatos marcantes dessa conflituosa e interessante relação entre o homem e o papilomavírus

Synthesis of folic acid-alliinase conjugate and its in situ inhibition of tumor cell proliferation by combination of alliin / 中国药科大学学报

@#To synthesize the folic acid-alliinase conjugate(FA-Alliinase), and to study its targeting and antitumor activity on cervical cancer HeLa cells. FA-Alliinase I and FA-Alliinase II were synthesized by two methods. The couping ratios of two conjugates measured were 12 and 31, respectively. The FA-Alliinase II with high coupling ratio was selected and its structure was characterized preliminarily. The activity of alliinase retained about 50% in FA-Alliinase II determined by HPLC. The specific effect of FA-Alliinase II on HeLa cells was observed by confocal laser and flow cytometry. The antitumor activity of conjugate combined with alliin was determined by MTT, and IC50 of alliin was(127. 6±2. 3)μmol/L. This study provides a direct evaluation method for the synthesis and optimization of FA-Alliinase.

Purification and identification of novel cytotoxic oligopeptides from soft coral Sarcophyton glaucum / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Globally, peptide-based anticancer therapies have drawn much attention. Marine organisms are a reservoir of anticancer peptides that await discovery. In this study, we aimed to identify cytotoxic oligopeptides from Sarcophyton glaucum. Peptides were purified from among the S. glaucum hydrolysates produced by alcalase, chymotrypsin, papain, and trypsin, guided by a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay on the human cervical cancer (HeLa) cell line for cytotoxicity evaluation. Purification techniques adopted were membrane ultrafiltration, gel filtration chromatography, solid phase extraction (SPE), and reversed-phase high-performance liquid chromatography (RP-HPLC). Purified peptides were identified by de novo peptide sequencing. From papain hydrolysate, three peptide sequences were identified: AGAPGG, AERQ, and RDTQ (428.45, 502.53, and 518.53 Da, respectively). Peptides synthesized from these sequences exhibited cytotoxicity on HeLa cells with median effect concentration (EC50) values of 8.6, 4.9, and 5.6 mmol/L, respectively, up to 5.8-fold stronger than the anticancer drug 5-fluorouracil. When tested at their respective EC50, AGAPGG, AERQ, and RDTQ showed only 16%, 25%, and 11% cytotoxicity to non-cancerous Hek293 cells, respectively. In conclusion, AERQ, AGAPGG, and RDTQ are promising candidates for future development as peptide-based anticancer drugs.

Analysis of Pharmacological Effect of Total Tannins from Spatholobi Caulis on Cervical Cancer HeLa Cells by 3D Microfluidic Chip / 中国实验方剂学杂志

Objective:To study on the chemical constituents of total tannins from Spatholobi Caulis,and to analyze the pharmacodynamics and mechanism of total tannins from Spatholobi Caulis on cervical cancer HeLa cells. Method:UPLC-Q-TOF/MS was used to qualitatively analyze the composition of total tannins from Spatholobi Caulis.The appropriate concentration and time of administration were screened by 3 dimensional(3D) microfluidic chip.Flow cytometry was used to determine the effect of total tannins from Spatholobi Caulis on the cell cycle and apoptosis of cervical cancer HeLa cells and analyzed by FlowJo v10.0.7 and ModFit LT 3.2 software.Enzyme-linked immunosorbent assay(ELISA) was used to determine the changes of vascular endothelial growth factor(VEGF)-A and Caspase-3 factors in cervical cancer HeLa cells supernatant treated with total tannins from Spatholobi Caulis. Result:Total of 15 components in total tannins from Spatholobi Caulis were identified or inferred.The low,medium and high dosages of total tannins from Spatholobi Caulis were 0.5,1.0, 2.0 g·L-1 and the best time of administration was 36 h.The proportions of early and late apoptosis of cervical cancer HeLa cells increased significantly in the apoptosis analysis after being treated by total tannins from Spatholobi Caulis.The DNA synthesis early phase(G0/G1 phase) of cervical cancer HeLa cells significantly increased,and the DNA synthesis phase(S phase) and the DNA synthesis late phase(G2/M phase) reduced.After being treated with total tannins from Spatholobi Caulis,the expression of VEGF-A in cervical cancer HeLa cells supernatant was significantly decreased and the expression of Caspase-3 was significantly increased. Conclusion:Spatholobi Caulis is rich in tannins,which can significantly inhibit the proliferation of cervical cancer HeLa cells and promote its apoptosis.This paper can provide the basis for further research of total tannins from Spatholobi Caulis.

Separation and Purification of Alhagi sparsifolia n -butanol Extract Monomeric Compounds and Study on Their Effects on Human Cervical Cancer HeLa Cells / 中国药房

OBJECTIVE： To separate and purify Alhagi sparsifolia n-butanol extract monomeric compounds， and to investigate its effects on the proliferation and metastasis of human cervical cancer HeLa cells. METHODS： The n-butanol extract was separated and purified by silica gel column， Sephadex LH-20 gel column and prep-HPLC. The structures of compounds were analyzed and identified according to physicochemical properties and spectrum （mass spectrum， hydrogen spectrum， carbon spectrum） data. Using human cervical cancer HeLa cells as objects， 5-FU as positive control， MTT assay was used to detect the inhibitory rate of HeLa cells pretreated with different doses of compounds （6.25， 12.5， 25， 50， 100， 200 μg/mL）； IC50 was calculated to screen active monomers. Scratch test was used to investigate the effects of above active monomers （all 50 μg/mL） on the migration ability of HeLa cells. Kim’s formula was used to evaluate the effects of 5-FU separately combined with above active monomers [（3.125+6.25），（6.25+12.5），（12.5+25），（25+50）μg/mL]. RESULTS： Six compounds were isolated from the n-butanol extract part of A. sparsifolia and identified as butin （Ⅰ）， 3′，4′，7-trihydroxyisoflavone （Ⅱ）， p-methoxyphenylacetic acid （Ⅲ）， 4-hydroxyacetophenone （Ⅳ）， aurantiamide acetate （Ⅴ）， protocatechualdehydea （Ⅵ）. Compared with blank control group， 5-FU and each compound （5-FU：6.25-200 μg/mL， compound Ⅰ： 12.5-200 μg/mL； compound Ⅱ： 25， 50， 200  μg/mL； compound Ⅲ： 6.25， 100， 200 μg/mL； compound Ⅳ： 50， 100， 200 μg/mL； compound Ⅴ： 12.5， 25， 200 μg/mL； compound Ⅵ： 6.25-200 μg/mL） could significantly increase the cell inhibition rate. IC50 of compound Ⅰ， Ⅴ， Ⅵ were decreased significantly （P＜0.05 or P＜0.01）， and those of compound Ⅰ and Ⅵ were lower relatively. The migration distance of cells in 5-FU and compound Ⅰ and Ⅵgroups were decreased significantly， compared with blank control group （P＜0.05 or P＜0.01）. 5-FU separately combined with compound Ⅰ and Ⅵ showed additive and enhanced inhibitory effects on the proliferation of HeLa cells （synergistic index＞0.9）. CONCLUSIONS： Compounds Ⅰ-Ⅵ are isolated from Alhagi for the first time. Butin and protocatechualdehydea are active monomers of its n-butanol extract part. Above two monomers can inhibit the proliferation and migration of human cervical cancer Hela cells， with strong inhibitory effect in vitro， and stronger inhibitory effect combined with 5-FU than any compound alone.