CircRNA_PLEKHM3 regulates epithelial mesenchymal transformation of cervical cancer cells through the miR-320/KLF4 axis / 安徽医科大学学报

Objective @#To investigate the role and mechanism of circular RNA containing pleck strin homology do main of Pleck strin homology domain family M member 3 ( circRNA_PLEKHM3) in regulating epithelial mesenchy mal transition (EMT) behavior in cervical cancer cells through the miR-320 and KLF4 .@*Methods @#The expression levels of circRNA_PLEKHM3 in cervical cancer cells Hela and CaSki were detected by real time quantitative PCR (qRT PCR) . RNA fluorescence in situ hybridization was used to determine the localization of circRNA_PLEKHM3 in human cervical cancer epithelial cells CaSki . Dual luciferase reporter gene experiments were conducted to inves tigate the targeting relationship between circRNA_PLEKHM3 and miR-320 , as well as the targeting relationship be tween miR-320 and KLF4 . CaSki cells were overexpressed with circRNA_PLEKHM3. Additionally, three groups were set up: overexpression of miR-320 on the basis of circRNA_PLEKHM3 overexpression , silencing of KLF4 on the basis of circRNA_PLEKHM3 overexpression , and silencing of KLF4 on the basis of miR-320 overexpression . qRT PCR was performed to detect the expression levels of miR-320 in CaSki . Western blot experiments were con ducted to determine the expression of KLF4 and EMT markers including E-cadherin , N-cadherin , Vimentin , MMP-2 , and MMP-9 in CaSki cells . Transwell assays were performed to measure cell migration and invasion .@*Results@#The expression of circRNA_PLEKHM3 decreased in Hela and CaSki cells (P < 0 05) , mainly localized in the cytoplasm . The dual luciferase reporter gene experiment demonstrated a targeting relationship between miR-320 and circRNA_PLEKHM3 , as well as between KLF4 and miR-320 . Overexpression of circRNA_PLEKHM3 inhibited the protein expression of miR-320 , Ncadherin , Vimentin , MMP-2 , and MMP-9 , up regulated the protein expression of E-cadherin, and reduced cell migration and invasion ( P < 0.05) . Overexpression of miR 320 or silencing of KLF4 on the basis of circRNA_PLEKHM3 overexpression both promoted the protein expression of miR-320, N-cad herin , Vimentin , MMP-2 , and MMP-9 , down regulated the protein expression of E-cadherin , and increased cell migration and invasion (P < 0.05) . However , silencing of KLF4 on the basis of miR-320 overexpression inhibited the protein expression of KLF4 , N-cadherin , Vimentin , MMP-2 , and MMP-9 , up regulated the protein expression of E-cadherin , and reduced cell migration and invasion ( P < 0.05) . @*Conclusion @#Overexpression of circRNA _PLEKHM3 regulates EMT in cervical cancer cells through the miR -320/KLF4 axis .

Mechanism of malignant behavior in cervical cancer cells based on NOD2-mediated AMPK/mTOR signaling pathway / 安徽医科大学学报

Objective To explore the mechanism of malignant behavior of cervical cancer(CC)cells based on AMP-activated protein kinase(AMPK)/rapamycin target protein(mTOR)signaling pathway mediated by nucleo-tide-binding oligomerization domain receptor 2(NOD2).Methods Bioinformatics analysis was performed to deter-mine the expression of NOD2 in CC tissue.Plasmids targeting NOD2(shNOD2)and shRNAs negative control(shNC),NOD2 overexpression(NOD2)and vectors(Vec)were transfected into CC cells.The effect of NOD2 on the growth of CC cells was determined by cell counting kit-8 assay,colony formation and Transwell cell invasion as-say.Transcriptome analysis was performed by high throughput RNA sequencing(RNA-Seq).Western blot was used to detect the expression of NOD2,AMPK/mTOR signaling pathway and autophagy protein in the cell line.24 female BALB/c nude mice were randomly divided into four groups,with 6 mice in each group:vector group(Vec group),NOD2 overexpression group(NOD2 group),shNC group and shNOD2 group.The distant metastasis mod-el was established in mice,and the fluorescence intensity of lung metastasis was monitored and the number of lung metastasis nodules was counted.Results On-line database analysis showed that the expression of NOD2 in CC tis-sues was significantly higher than that in normal tissues,and there were significant differences in the mRNA expres-sion of NOD2 in different stages of CC(P＜0.05).In addition,the high expression of NOD2 was associated with poor overall survival and disease-free survival(P＜0.05).NOD2 overexpression promoted the proliferation,colony formation,migration and invasion of CC cells,while NOD2 knock-down was the opposite.Consistent with the re-sults in vitro,the lung colonization and lung metastasis of CC cells in NOD2 group were significantly higher than those in Vec group(P＜0.05),while those in shNOD2 group were significantly lower than those in shNC group(P＜0.05).RNA-Seq results showed that the expression of NOD2 was significantly related to AMPK signal activa-tion,mTOR signal inhibition,autophagy regulation pathway activation and autophagy formation.Compared with shNC group,the expression of phosphorylated AMPK and LC3 protein decreased significantly in shNOD2 group(P＜0.05),and the expression levels of phosphorylated mTOR and p62 protein increased significantly(P＜0.05).Compared with Vec group,the expression levels of LC3 and AMPK protein in NOD2 group increased significantly(P＜0.05),and the expression levels of phosphorylated mTOR and p62 protein decreased significantly(P＜0.05).Compared with shNC group,the point accumulation of GFP-mRFP-LC3 in shNOD2 group decreased signifi-cantly(P＜0.05).Compared with Vec group,the point accumulation of GFP-mRFP-LC3 increased significantly(P＜0.05).Conclusion NOD2 may promote the proliferation,migration and invasion of CC through AMPK/mTOR signal,and its mechanism partly involves autophagy activation.

Effects of ligustilide on chemotherapy resistance of cervical cancer cells / 中国药房

OBJECTIVE To investigate the effects of ligustilide on chemotherapy resistance of cervical cancer cells based on Hippo-Yes-associated protein （YAP） signaling pathway. METHODS Human cervical cancer cisplatin-resistant cells HeLa/DDP were divided into control group， cisplatin group （10 μmol/L cisplatin）， cisplatin+ligustilide low-， medium- and high-concentration groups （10 μmol/L cisplatin+25， 50， 100 μmol/L ligustilide）. The proliferation， apoptosis， migration and invasion of HeLa/DDP cells were all detected in each group. The mRNA expressions of YAP and transcriptional coactivator with PDZ binding motif （TAZ） as well as the protein expressions of YAP， TAZ， matrix metalloproteinase 2 （MMP2）， Ki67， cleaved-caspase-3 and caspase-3 were determined in HeLa/DDP cells. RESULTS Compared with control group， the inhibitory rate， apoptotic rate and cleaved- caspase-3/caspase-3 of cisplatin group were increased significantly； scratch healing rate， the number of invasive cells， the mRNA expressions of YAP and TAZ， and the protein expressions of YAP， TAZ， MMP2 and Ki67 were decreased significantly in cisplatin group （P＜0.05）. Compared with cisplatin group， the inhibitory rate of cell proliferation， apoptotic rate and cleaved-caspase-3/ caspase-3 were further increased in cisplatin+ligustilide low-， medium- and high-concentration groups， while scratch healing rate， the number of invasive cells， the mRNA expressions of YAP and TAZ， and the protein expressions of YAP， TAZ， MMP2 and Ki67 were further decreased， in a dose-dependent manner （P＜0.05）. CONCLUSIONS Ligustilide can increase the sensitivity of drug-resistant cervical cancer cells to cisplatin by inhibiting Hippo-YAP signaling pathway.

Phytochemical profile and biological activities of Bromelia antiacantha extracts

Abstract Reports from popular medicine usually act as a basis for the development of new drugs from natural compounds with therapeutic actions for serious diseases and prevalence such as cancer. Bromelia antiacantha Bertol. is a species of the Bromeliaceae family, considered an unconventional food plant, found in the south and midwest regions of Brazil. Despite the high nutritional content and pharmacological potential of its fruits, few scientific studies report its biological actions. Thus, this study evaluates the phytochemical profile of aqueous and ethanol extracts obtained from B. antiacantha fruits, as well as their possible antioxidant, antitumor, and cytotoxic activities. The aqueous extract exhibited phenolic compounds and flavonoids, while ethanol extracts indicated the presence of flavonoids and coumarin in their composition, regardless of the region of collection. The ethanolic extract demonstrated a more promising antioxidant effect than the aqueous extract and also induced a significant inhibition in the viability of human cervical cancer cells of the SiHa strain. In addition, treatment with both extracts did not alter the viability of non-tumor cells of the immortalized human keratinocyte lineage (HaCaT). These results bring new data about extracts obtained from a native plant, edible and traditionally used in popular medicine, opening new perspectives for its possible therapeutic application.

Resumo Relatos da medicina popular costumam atuar como referencial para o desenvolvimento de novos fármacos a partir de moléculas naturais com ações terapêuticas para doenças de alta gravidade e prevalência como o câncer. Bromelia antiacantha Bertol. é uma espécie da família Bromeliaceae, considerada uma planta alimentícia não convencional (PANC), encontrada nas regiões sul e centro-oeste do Brasil. Apesar do alto teor nutritivo e potencial farmacológico de seus frutos, poucos estudos científicos relatam suas ações biológicas. Desta forma, este estudo avalia o perfil fitoquímico de extratos aquoso e etanólico obtidos de frutos de B. antiacantha, bem como a sua possível ação antioxidante, antitumoral e citotóxica. O extrato aquoso apresentou compostos fenólicos e flavonoides, enquanto os extratos etanólicos apontam a presença de flavonóides e cumarina em sua composição, independente da região de coleta. O extrato etanólico demonstrou efeito antioxidante mais promissor do que o extrato aquoso e também induziu uma inibição significativa na viabilidade de células humanas de câncer cervical da linhagem SiHa. Além disso, o tratamento com ambos extratos não alterou a viabilidade de células não tumorais da linhagem de queratinócitos humanos imortalizados (HaCaT). Estes dados trazem novas informações sobre extratos obtidos de uma espécie vegetal nativa, comestível e já utilizada tradicionalmente, mas abrindo novas perspectivas quanto a possíveis aplicações terapêuticas.

Phytochemical profile and biological activities of Bromelia antiacantha extracts / Perfil fitoquímico e atividades biológicas de extratos de Bromelia antiacantha

Reports from popular medicine usually act as a basis for the development of new drugs from natural compounds with therapeutic actions for serious diseases and prevalence such as cancer. Bromelia antiacantha Bertol. is a species of the Bromeliaceae family, considered an unconventional food plant, found in the south and midwest regions of Brazil. Despite the high nutritional content and pharmacological potential of its fruits, few scientific studies report its biological actions. Thus, this study evaluates the phytochemical profile of aqueous and ethanol extracts obtained from B. antiacantha fruits, as well as their possible antioxidant, antitumor, and cytotoxic activities. The aqueous extract exhibited phenolic compounds and flavonoids, while ethanol extracts indicated the presence of flavonoids and coumarin in their composition, regardless of the region of collection. The ethanolic extract demonstrated a more promising antioxidant effect than the aqueous extract and also induced a significant inhibition in the viability of human cervical cancer cells of the SiHa strain. In addition, treatment with both extracts did not alter the viability of non-tumor cells of the immortalized human keratinocyte lineage (HaCaT). These results bring new data about extracts obtained from a native plant, edible and traditionally used in popular medicine, opening new perspectives for its possible therapeutic application.

Relatos da medicina popular costumam atuar como referencial para o desenvolvimento de novos fármacos a partir de moléculas naturais com ações terapêuticas para doenças de alta gravidade e prevalência como o câncer. Bromelia antiacantha Bertol. é uma espécie da família Bromeliaceae, considerada uma planta alimentícia não convencional (PANC), encontrada nas regiões sul e centro-oeste do Brasil. Apesar do alto teor nutritivo e potencial farmacológico de seus frutos, poucos estudos científicos relatam suas ações biológicas. Desta forma, este estudo avalia o perfil fitoquímico de extratos aquoso e etanólico obtidos de frutos de B. antiacantha, bem como a sua possível ação antioxidante, antitumoral e citotóxica. O extrato aquoso apresentou compostos fenólicos e flavonoides, enquanto os extratos etanólicos apontam a presença de flavonóides e cumarina em sua composição, independente da região de coleta. O extrato etanólico demonstrou efeito antioxidante mais promissor do que o extrato aquoso e também induziu uma inibição significativa na viabilidade de células humanas de câncer cervical da linhagem SiHa. Além disso, o tratamento com ambos extratos não alterou a viabilidade de células não tumorais da linhagem de queratinócitos humanos imortalizados (HaCaT). Estes dados trazem novas informações sobre extratos obtidos de uma espécie vegetal nativa, comestível e já utilizada tradicionalmente, mas abrindo novas perspectivas quanto a possíveis aplicações terapêuticas.

LncRNA PCAT-1 Reduces Sensitivity of Cervical Carcinoma Cells to Cisplatin by Regulating STAT3 and PTEN Expression / 肿瘤防治研究

Objective To investigate the role and regulatory mechanism of lncRNAs PCAT-1 in the sensitivity of cervical cancer cells to DDP. Methods The expressions of PCAT-1 in human cervical cancer cell lines (HeLa and SiHa) and DDP-resistant cell lines (HeLa/DDP and SiHa/DDP) were analyzed by real-time PCR. After PCAT-1 silencing and overexpression in HeLa/DDP and SiHa/DDP cells, CCK-8 and flow cytometry were used to detect cell viability ability and cell cycle, respectively. Western blot was used to detect the protein expression of STAT3 and PTEN. Results The DDP resistance index of HeLa/DDP cells to HeLa cells was 4.49, while that of SiHa/DDP cells to SiHa cells was 6.87. The expression levels of PCAT-1 in HeLa/DDP and SiHa/DDP cells were significantly higher than those in HeLa and SiHa cells, respectively (P < 0.05). The overexpression of PCAT-1 reduced the sensitivity of HeLa/DDP and SiHa/DDP cells to DDP, enhanced the proportion of S phase in cell cycle, and decreased the proportion of G0-G1 and G2-M phases (P < 0.05). The silencing of PCAT-1 increased the sensitivity of HeLa/DDP and SiHa/DDP cells to DDP, decreased the proportion of S phase in the cell cycle, and enhanced the proportion of G0-G1 and G2-M phase (P < 0.05). Overexpression of PCAT-1 promoted STAT3 protein expression but inhibited PTEN protein expression in HeLa/DDP and SiHa/DDP cells (P < 0.05). The silencing of PCAT-1 inhibited STAT3 protein expression but promoted PTEN protein expression in HeLa/DDP and SiHa/DDP cells (P < 0.05). Conclusion PCAT-1 is upregulated in HeLa/DDP and SiHa/DDP cells. PCAT-1 reduces the sensitivity of HeLa/DDP and SiHa/DDP cells to DDP by upregulating the expression of STAT3 and downregulating the expression of PTEN.

A study on the mechanism of mi-362 targeting six1 inhibiting proliferation and migration of cervical cancer cells / 实用肿瘤学杂志

Objective To investigate the mechanism of Mi-362 targeting Six1 inhibiting proliferation and migration of cer-vical cancer cells. Methods The expression levels of microRNA-362 in cervical cancer tissues and adjacent tissues were measured in 142 patients with cervical cancer in our hospital. At the same time,Hela cancer cell group,microRNA-362 mimics group and mi-croRNA-362 inhibitor group were set up to determine the viability of cancer cells,the number of monoclonal formation of cancer cells,the apoptotic rate of cancer cells,cell cycle,the number of perforations,and the levels of microRNA-362 and Six1 in cervical cancer fluid of Hela. Results The expression level of miR-362 in cervical cancer tissue was lower than that in adjacent tissue(P<0. 05). The higher the infiltration of lymphatic vessel space,pathological stage,TNM stage,lymph node metastasis and depth of infil-tration,the lower the expression rate of miR-362(P<0. 05). The OD value and survival rate in the miR-362 mimics group were lower than those in the Hela cancer cells group(P<0. 05),while the OD value and survival rate in the mir-362 inhibitor group were higher than those in the Hela cancer cells group and the miR-362 mimics group(P<0. 05). The number of clones formed in the miR-362 mimics group was lower than that in the Hela cancer cell group(P<0. 05),and the number of clones formed in the miR-362 inhibitor group was higher than that in the Hela cancer cell group and the miR-362 mimimics group(P<0. 05). The apoptotic rate of miR-362 mimics group was higher than that of Hela cancer cell group(P<0. 05),and that of miR-362 inhibitor group was lower than that of Hela cancer cell group and miR-362 mimics group(P<0. 05). The G1 phase in the miR-362 mimics group was higher than that in the Hela cancer cell group(P<0. 05),and the G1 phase in the miR-362 inhibitor group was lower than that in the Hela cancer cell group and the miR-362 mimimics group(P<0. 05). The number of cell membrane penetration in the miR-362 mimics group was lower than that in the Hela cancer group(P < 0. 05),and that in the miR-362 inhibitor group was higher than that in the Hela cancer group and the miR-362 mimimics group(P<0. 05). The expression level of miR-362 in the miR-362 mimics group was higher than that in the Hela cancer cell group(P<0. 05),and the expression level of miR-362 in the miR-362 inhibitor group was lower than that in the Hela cancer cell group and the miR-362 mimics group(P<0. 05). The expression level of Six1 in the miR-362 mimics group was lower than that in the Hela cancer cell group(P<0. 05),and that in the miR-362 inhibitor group was higher than that in the Hela cancer cell group and the miR-362 mimics group(P<0. 05). Conclusion miR-362 plays an important inhibition in the occurrence and development of cervical cancer,and its mechanism is related to the inhibition of prolifer-ation,migration and invasion of cancer cells by microRNA-362 through negative regulation of Six1.

Inhibiting expression of polyamines regulator-1 can enhance the antitumor activity of dexamethasone on human cervical cancer cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the influence of inhibiting expression of polyamine-modulated factor (PMF-1) on the antitumor effect of glucocorticoid dexamethasone (DEX) in human cervical cancer Caski cells. Methods: siRNAs which target human PMF-1 gene were designed and synthesized, and their effect on the expression of PMF-1 in Caski cells was evaluated by Western blotting. The PMF-1 down-regulated and control Caski cells were treated with DEX, and then the affect of PMF-1 down regulation on the sensitivity of the tumor cells to DEX was analyzed. MTT method was used to detect cell proliferation, flow cytometry was used to analyze cell cycle, Western blotting method was used to evaluate expression level of glucocorticoids receptor (GR), and HPLC was used to analyze intracellular polyamine content. Results: The transient transfection of Caski cells with siRNAwhich targets PMF-1 gene can significantly reduce the expression level of PMF-1 protein. Compared with the control cells, treating PMF-1 down-regulated Caski cells with DEX can more effectively inhibit cell proliferation（P<0.01), up regulate GR expression, arrest cell cycle at G2 stage（P<0.01), and also significantly reduce intracellular polyamine level（P<0.01). Conclusion：Inhibiting PMF-1 expression can enhance antitumor pharmacological activity of DEX against human cervical cancer cells, and the underlying mechanism may be related with enhanced cell cycle inhibition and decreased intracellular polyamine level.

TMPYP4 exerted antitumor effects in human cervical cancer cells through activation of p38 mitogen-activated protein kinase

BACKGROUND: The aim of the present study was to investigate the potential effects of the 5,10,15,20-tetrakis (1-methylpyridinium-4-yl) porphyrin (TMPyP4) on the proliferation and apoptosis of human cervical cancer cells and the underlying mechanisms by which TMPyP4 exerted its actions. RESULTS: After human cervical cancer cells were treated with different doses of TMPyP4, cell viability was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method, the apoptosis was observed by flow cytometry (FCM), and the expression of p38 mitogen-activated protein kinase (MAPK), phosphated p38 MAPK (p-p38 MAPK), capase-3, MAPKAPK2 (MK-2) and poly ADP-ribose polymerase (PARP) was measured by Western blot analysis. The analysis revealed that TMPyP4 potently suppressed cell viability and induced the apoptosis of human cervical cancer cells in a dose-dependent manner. In addition, the up-regulation of p-p38 MAPK expression levels was detected in TMPyP4-treated human cervical cancer cells. However, followed by the block of p38 MAPK signaling pathway using the inhibitor SB203580, the effects of TMPyP4 on proliferation and apoptosis of human cervical cancer cells were significantly changed. CONCLUSIONS: It was indicated that TMPyP4-inhibited proliferation and -induced apoptosis in human cervical cancer cells was accompanied by activating the p38 MAPK signaling pathway. Taken together, our study demonstrates that TMPyP4 may represent a potential therapeutic method for the treatment of cervical carcinoma.

The regulatory effect of 5-aza-2,-deoxycytidine in cervical cancer cells / 实用医学杂志

Objective To discuss the regulatory effect of 5-aza-2 ,-deoxycytidine on P16 and MGMT in cervical cancer cells. Methods After four kinds of cervical cancer cells (HeLa, SiHa, C33A and CaSki) were treated with 5-Aza-dC , MSP was used to detect the methylation variation of P16 and MGMT , and fluorogenic quantitative PCR and Western blot were employed for determination of P16 and MGMT expression. MTT and Annexin V-FITC/PI double staining were adopted for detection of cell proliferation and apoptosis. Results Both P16 and MGMT exhibited methylation in four kinds of cervical cancer cells , and after treatment with 5-Aza-dC ,their methylation levels were reversed. 5-Aza-dC was able to inhibit p16 and MGMT expression in the cervical cancer cells, and can also suppress cell proliferation and promote apoptosis. Conclusions Although methylation of P16 and MGMT are present in cervical cancer cells, their expression level was still high. Therefore, regulation of P16 and MGMT expression may be affected by other factors. 5-Aza-dC can suppress the growth of cervical cancer cells. Although 5-Aza-dC reverse the methylation levels of P16 and MGMT, it inhibits their gene expression. More experiments are needed to verify the hidden reasons and mechanisms.

Effects of autophagy on the proliferation and migration of cervical cancer cells / 实用肿瘤学杂志

Objective To investigate the effect of autophagy on the proliferation and migration of cervi-cal cancer cells ,as well as the underlining mechanisms .Methods Rapamycin was used to induce the autophagy in HeLa cells,formation of autophagosomes was observed by staining with acridine orange under fluorescence mi -croscope.Western blot was used to detect the expression of LC 3 and PI3K/Akt/mTOR in HeLa cells.The LC3 plasmid was transfected into HeLa cells .The distribution of LC3 in cells and the expression of LC3 was identified by fluorescence microscope and Western blot ,respectively.The autophagy was inhibited with 3-methyladenine(3-MA )in HeLa cells.The cell proliferation was monitored by RTCA real -time instrument.Transwell chamber was carried out to assess cell migration .Results After 6h of rapamycin treatment ,the expression of LC3B was in-creased in HeLa cells ( P<0 .05 ) .The proliferative and migration ability were weakened compared to wild type HeLa cells(P<0.05).The same results in the presence of 3-MA.The expression of PI3K/AKT/mTOR path-way proteins were activated by rapamycin treatment and LC 3 overexpression(P<0.05).Conclusion Autophagy can suppress the proliferation and migration in cervical cancer cells ,which may relate to PI3K/Akt/mTOR path-way.

Dosimetry study on photodynamic effect of PSD-007 on human cervical cancer cell line Hela / 国际生物医学工程杂志

Objective To investigate the photochemotherapeutic effect and the main affecting factors of PSD-007 on human cervical cancer Hela in vitro.Methods Hela cells were treated with different concentrations of PSD-007 (0,3.125,6.25,12.5,25,50,100 μg/ml) for 2 h under the influence of low-level laser (635 nm) therapy at different doses (0,0.6,1.2,2.4,4.8,9.6 J/cm2).Then the OD values and survival rates of Hela cells were measured by MTT assay compared with breast cancer cells MCF-7 in same treatment.Hela cells were treated with 12.5 μg/ml of PSD-007 for 2 h and were treated with different intensities of laser (1.2,2.4,4.8 J/cm2).The cellular apoptosis rate and cell cycle phase distribution of Hela were measured by a flow cytometry (FCM).Results Survival rates of Hela cells declined with more than 25 μg/ml of PSD-007 only,and significant difference in the inhibitory between the PDT group and control group was observed (P＜0.05).The survival rates of Hela after PDT was decreased by the concentration of sensitizer and dose of laser.There were no significant differences of cell survival rates among the groups with concentrations more than 12.5 μg/ml and laser energy density more than 4.8 J/cm2.The FCM assay showed a G0/G1 cell cycle arrest in a time-dependent manner.Conclusions PSD-007 has a photodynamic effect on Hela in vitro.Photodynamic effect of PSD-007 was more significant in Hela than MCF-7.Less photosensitizer and laser energy density were needed.

Influence of folate on fragile histidine triad gene expression,cell proliferation and apoptosis in cervical cancer cell lines / 中华流行病学杂志

Objective To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation,expression of mRNA and protein of fragile histidine triad(FHIT)gene in cervical cancer cells. Methods Cervical cancer cell lines including CaSki(HPV16-positive)and C33A(HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR(MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR,respectively. Results Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r=0.98,P<0.001;CaSki:r=0.98,P<0.001) and the apoptosis rate increased(C33A:r=0.98,P<0.001;CaSki:r=0.99,P<0.001)along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/ml in both C33A and CaSki cells. Comparing with the control group,the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells,and showing that the FHIT gene mRNA expression(C33A:r=0.96,P<0.001;CaSki:r=0.94,P<0.001)and protein expression (C33A:r=0.96,P<0.001;CaSki:r=0.97,P<0.001) both increased along with the increase of folate concentration. Conclusion Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,so would reverse the aberration mRNA and protein expression of FHIT gene.

Influence of folate on fragile histidine triad gene expression,cell proliferation and apoptosis in cervical cancer cell lines / 中华流行病学杂志

Objective To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation,expression of mRNA and protein of fragile histidine triad(FHIT)gene in cervical cancer cells. Methods Cervical cancer cell lines including CaSki(HPV16-positive)and C33A(HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR(MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR,respectively. Results Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r=0.98,P<0.001;CaSki:r=0.98,P<0.001) and the apoptosis rate increased(C33A:r=0.98,P<0.001;CaSki:r=0.99,P<0.001)along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/ml in both C33A and CaSki cells. Comparing with the control group,the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells,and showing that the FHIT gene mRNA expression(C33A:r=0.96,P<0.001;CaSki:r=0.94,P<0.001)and protein expression (C33A:r=0.96,P<0.001;CaSki:r=0.97,P<0.001) both increased along with the increase of folate concentration. Conclusion Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,so would reverse the aberration mRNA and protein expression of FHIT gene.

Significance of aberrant expression of DNA methyltransferase 1 in cervical cancerization tissue and cervical cancer cell lines / 肿瘤研究与临床

Objective To explore the effects of aberrant expression of DNA methyltransferase 1 (DNMT1) in cervical cancerization tissue and cervical cancer cells.Methods Cervical tissues were collected from 80 cases with a diagnosis of invasive cervix squamous cell carcinoma (SCC),53 cases with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),52 cases with low-grade cervical intraepithelial neoplasia (CIN Ⅰ)and 53 cases with normal cervix (NC).Meanwhile,Caski (HPV16-positive) and C33A (HPV-negative) cells selected from cervical cancer cell lines were cultured routinely in vitro.The expression of DNMT1 protein and mRNA were examined by Western blot analysis and real-time PCR (qRT-PCR) in the tissues and cells,respectively.Results The levels of DNMT1 protein were 1.33,1.84 and 2.28,and the Ct-ratios (DNMT1/β-actin) of DNMT1 mRNA were 1.27,1.27 and 1.26 in CIN Ⅰ,CIN Ⅱ/Ⅲand SCC group,respectively.Comparing with NC group,the expression of DNMT1 protein or mRNA was elevated in deficient cervical groups,with statistical significance (F =110.57,P ＜ 0.001,F =2.68,P =0.048).The expression levels of DNMT1 protein were increased steadily according to severity of the cervix lesions (x2tend =50.80,P ＜ 0.001),however,the expression of DNMT1 mRNA was not observed the same tendency (x2tend =3.63,P ＞ 0.05).The results from experiment in vitro showed that the levels of DNMT1 protein or mRNA were both higher in Caski cell than in C33A cell,especially for DNMT1 mRNA with significantly difference (t =7.134,P =0.002).Conclusion Aberrant expression of DNMT1 protein or mRNA could link with the risk of cervical cancerization by both transcriptional and posttranscriptional mechanisms.There would be a synergistic effect between overexpression of DNMT1 and HPV16 infection in the progression of cervix carcinogenesis.

Radiation sensitization by dihydroartemisinin on human HeLa cells of cervical cancer / 中华放射医学与防护杂志

Objective To investigate the radiosensitizing effects of dihydroartemisinin(DHA)on human HeLa cells of cervical cancer irradiated by X rays.Methods Cell growth kinetics was determined using MTF assay.Cell survival was analyzed by elonogenic assay.The change of cell cycle and apeptosis was measured by flow cytometry.Results Dihydroartemisinin inhibited the growth of HeLa cells of human cervical cancer and showed a dose-dependent and time-dependent manner.Dihydroartemisinin(20 μmol/L)showed the radiosensitizing effects on HeLa cells,and the sensitizing enhancement ratio(SER)was 1.47.Dihydroartemisinin abrogated radiation-induced G2 arrest of the tested HeLa cells,the G2 ratio of medicine + radiation group dechned from 73.58% to 48.31%.Dihydroartemisinin enhanced the apoptosis of HeLa cells by X-irradiation,the apoptosis rates of medicine + radiation group significantly increased from 29.46%,48.04%,70.21% to 45.79%,66.36% and 79.58%,respectively for 2,4 and 6 Gy.Conclusions Dihydroartemisinin could increase the radiesensitivity of HeLa cells of human cervical cancer.Abrogation of radiation-induced C2 arrest could be part of the mechanism.

Adenosine 5'-triphosphate induced NF-kappa B and intracellular calcium mobilization in cervical cancer cells / 대한산부인과학회지

OBJECTIVE: To know the effect of adenosine 5'-triphosphate (ATP) on intracellular calcium level and cell proliferation in cervical cancer cells. METHODS: Study design: Four different human cervical cancer cell lines (Caski, C33A, HeLaS3 and SiHa) were used in this study. The change of intracellular calcium level, cell proliferation and the activity of proliferation- and calcium-related transcription factors by extracellular ATP were examined in these cell lines. RESULTS: Extracellular ATP induced calcium mobilization, cell proliferation and the activation of NF-kappa B in all cell lines used. CONCLUSION: These results suggest that calcium mobilization and NF-kappa B dependent signaling pathway play an important role in the cell proliferation by ATP in cervical cancer.