The effect and mechanism of miR-199a-5p on the radiosensitivity of cervical cancer CaSki cells / 中国医师杂志

Objective:To investigate the effect and potential mechanism of miR-199a-5p on the radiosensitivity of cervical cancer CaSki cells.Methods:Cervical cancer CaSki cells were cultured in vitro. MiR-199a-5p mimics (miR-199a-5p mimics) were transfected into cervical cancer CaSki cells (miR-199a-5p group) with liposome by Lipofectamine 2000. CaSki cells transfected with mimics control were used as negative control (NC group) and non transfected CaSki cells were used as blank control (control group). After X-ray irradiation, real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-199a-5p in cells of each group. The effects of miR-199a-5p on radiosensitivity and apoptosis of CaSki cells were detected by clone formation assay and flow cytometry apoptosis assay. Bioinformatics software was used to predict the target gene of miR-199a-5p. Double luciferase reporter gene assay and Western blot were used to verify the targeting relationship between miR-199a-5p and thyroid hormone receptor interaction factor 4 (TRIP4). Results:The expression of miR-199a-5p in miR-199a-5p group was significantly higher than that in control group ( P<0.05). After X-ray irradiation, the expression of miR-199a-5p was more obvious ( P<0.05); Overexpression of miR-199a-5p could reduce the clonogenic ability and promote the apoptosis of CaSki cells ( P<0.05); Overexpression of miR-199a-5p could further reduce the clonal formation and promote the apoptosis of irradiated cells ( P<0.05); Double luciferase reporter gene experiment and Western blot confirmed that miR-199a-5p could target and negatively regulate TRIP4. Conclusions:miR-199a-5p can increase the radiosensitivity of cervical cancer CaSki cells by negatively regulating the expression of TRIP4.

Effect of lncRNA CCAT2 on proliferation and cell cycle of cervical cancer CaSki cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of long non-coding RNA (lncRNA)-CCAT2 on the proliferation and cell cycle of cervical cancer cells. Methods: The expression of CCAT2 in 3 cervical cancer cell lines (HeLa, C-33A, and CaSki) was detected by qPCR and the cell line with the highest expression level was selected for subsequent experiments. CCAT2 overexpression and interference vectors were designed and synthesized. After transfection, qPCR was performed to detect the transfection efficiency. The cells were divided into 5 groups: control, sh-EV (empty vector), overExp-EV , sh-CCAT2, and overExp-CCAT2. MTT assay was performed to evaluate cell viability. Flow cytometry was performed to measure cell cycle. WB was performed to detect the expressions of Ki67, cyclin D1, and cyclin dependent kinase 4 (CDK4). Results: Among HeLa, C-33A, and CaSki cells, the highest expression of CCAT2 was found in CaSki cells. CCAT2 overexpression and interference vectors were successfully transfected into the CaSki cells. Compared with the control group, the cells viability and proliferation in the sh-CCAT2 group was significantly decreased (all P<0.01), the proportion of cells in the G1 phase was significantly increased (P<0.01), and the expression levels of Ki67, cyclin D1, and CDK4 were significantly decreased (all P<0.01). However, in the overExp-CCAT2 group, the cell proliferation was enhanced and the expression levels of Ki67, cyclin D1, and CDK4 were significantly increased (all P<0.01). Conclusion: CCAT2 affects proliferation and cell cycle of cervical cancer cells by regulating the expressions of their associated proteins.

C-phycocyanin inhibits epithelial-mesenchymal transformation ofTGF-β1-induced cervical cancer Caski cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of C-phycocyanin (C-PC) on the epithelial-mesenchymal transition (EMT) of cervical cancer Caski cells induced by transforming growth factor beta1 (TGF-β1). Methods: According to different treatment methods, Caski cells were divided into three groups: 10 ng/ml TGF-β1 treatment group, 10 ng/ml TGF-β1+300 μg/ml C-PC co-treatment group and control group (untreated). After 24 h of treatment, the morphological changes of Caski cells were observed, and the effects of TGF-β1 and C-PC on the migration and invasion of Caski cells were detected by Scratch test and Transwell test, respectively. Western blotting was used to detect the effect of C-PC on the expression of epithelial phenotypic marker protein E-cadherin and stromal phenotypic marker protein N-cadherin in TGF-β1-induced Caski cells, and qPCR was used to detect the mRNA expressions of EMT related factors Snail, Zeb1 and Twist. Results: Caski cells in the TGF-β1 treatment group lost the characteristics of the original epithelial phenotype, while the cells in the TGF-β1+C-PC co-treatment group maintained the characteristics of normal epithelial phenotype; the migration rate ([60.0±1.4]% vs [33.5±2.2]%, [40.0±2.8]%, both P<0.05) and the number of invasive transmembrane cells ([108.2±6.2] vs [25.2±3.1], [39.8±5.4], both P<0.01]) of Caski cells in the TGF- β1 treatment group were significantly higher than those in the co-treatment group and the control group. Compared with the control group, the expression of E-cadherin in Caski cells treated with TGF-β1 decreased significantly (P<0.05), while the mRNA expressions of Twist, Snail and Zeb1 increased significantly (all P<0.05); However, co-treatment with C-PC reversed above changes (P<0.05 or P<0.01), and significantly decreased the protein expression level of N-cadherin (P< 0.05). Conclusion: C-PC treatment can inhibit the invasion and metastasis ability of Caski cells induced by TGF-β1 and further affects the EMT process. The mechanism may be related to the decrease of mRNAexpressions of Twist, Snail and Zeb1 by C-PC treatment. ·

miR-140 inhibits PD-L1 expression to enhance sensitivity of cervical cancer HeLa and Caski cells to oxaliplatin / 中国肿瘤生物治疗杂志

@# Objective: To investigate whether miR-140 could increase the sensitivity of cervical cancer (CC) to oxaliplatin by downregulating the expression of programmed death-1 (PD-L1). Methods: qPCR was used to analyze miR-140 expression in normal human cervical cells, CC cells and oxaliplatin-resistant CC cells. Cells were transfected with miR-140 mimic, and then, the proliferation of CC cells and oxaliplatin-resistant CC cells was detected by using CCK-8 assay, and the colony formation rate of CC cells was obtained by using colony formation assay. Starbase and TargetScan were used to predict the targeted binding site of miR-140 and PD-L1, and the influence of miR-140 on the expression of PD-L1 was validated by dual luciferase reporter gene assay.Annexin V FITC/PI double staining and Wb assays were used to detect the effect of over-expression of miR-140 or both over-expression of PD-L1 and miR140 on the apoptosis, migration and expression of apoptosis-related proteins in CC cells after treatment with oxaliplatin. Moreover, transplantation tumor of CC cell lines was established in nude mice to assess the effects of miR-140 on enhancing the sensitivity of tumors to oxaliplatin. Results: The expression of miR-140 was significantly decreased in oxaliplatin-resistant CC cells (P<0.01). Over-expression of miR140 could significantly increase the sensitivity of oxaliplatin-resistant CC cells to oxaliplatin (P<0.05), and inhibit the CC cells proliferation and colony formation (P<0.01). miR-140 showed targeted binding to PD-L1 3'-UTR and inhibited its expression. Over-expression of miR-140 significantly promoted CC cell migration and apoptosis (P<0.01). However, co-transfection of PD-L1 counteracts the effects of miR-140 on cell metastasis and apoptosis (all P<0.05). In addition, xenograft tumor model in mice also verified that miR-140 could promote the sensitivity of tumors to oxaliplatin. Conclusion: miR-140 increases the sensitivity of CC to oxaliplatin through inhibition of PD-L1 expression. Therefore, up-regulation of miR-140 or down-regulation of PD-L1 in combination with oxaliplatin may be a novel strategy for the treatment of Oxaliplatin-resistant CC.

Effects of genistein on anti-tumor activity of cisplatin in human cervical cancer cell lines

OBJECTIVE: To investigate the effect of genistein on the anticancer effects of chemotherapeutic agents, we examined the effect of a genistein and cisplatin combination on CaSki human cervical cancer cells. METHODS: After the cervical cancer cells (HeLa cells, CaSki cells) had been cultured, cisplatin and genistein were added to the culture medium, and the cell activity was measured using MTT assay. The CaSki cells were cultured in a medium containing cisplatin and genistein, and then, the cells were collected in order to measure p53, Bcl2, ERK, and caspase 3 levels by western blotting. RESULTS: Both the HeLa and CaSki cells had decreased cell viabilities when the cisplatin concentration was 10 μM or higher. When combined with genistein, the cell viabilities of the HeLa and CaSki cells decreased at cisplatin concentrations of 8 μM and 6 μM, respectively. The administration of genistein increased the toxicity of cisplatin in the HeLa and CaSki cells. In the CaSki cells, the p-ERK1/2 level decreased by 37%, the p53 expression level increased by 304%, and the cleaved caspase 3 level increased by 115% in the cisplatin+genistein group compared to that in the cisplatin group. Bcl2 expression was reduced by 69% in the cisplatin+genistein group compared to that in the cisplatin group. CONCLUSION: Genistein enhances the anticancer effect of cisplatin in CaSki cells, and can be used as a chemotherapeutic adjuvant to increase the activity of a chemotherapeutic agent.

Effects of lobaplatin on proliferation and invasion of cervical cancer CaSki cells / 国际肿瘤学杂志

Objective To investigate the effects of lobaplatin on proliferation and invasion of cervical cancer CaSki cells.Methods Human cervical cancer CaSki cells were randomly divided into blank control group,2,6 and 12 μg/ml lobaplatin groups by random number table method.The proliferations of the cells were detected by methyl thiazolyl tetrazolium (MTT).The morphological changes of the cells were observed by inverted microscope.The invasive abilities of the cells were detected by Transwell invasion test.The protein expressions of extracellular signal-regulated kinase (ERK) and phospho-extracellular signal-regulated kinase (p-ERK) were detected by Western blotting.Results The absorbance (A) values of blank group,2,6,12 μg/ml lobaplatin groups cultured for 24 h were 0.513 ± 0.023,0.428 ± 0.014,0.380 ± 0.012 and 0.300 ± 0.013 respectively,those of the cells cultured for 48 h were 0.831 ± 0.024,0.558 ± 0.019,0.415 ± 0.015 and 0.088 ±0.009 respectively,and those of the cells cultured for 72 h were 1.153 ±0.022,0.572 ± 0.023,0.201 ± 0.017 and 0.052 ± 0.014 respectively.The differences were statistically significant (F =12.922,P ＜ 0.001;F =10.192,P ＜ 0.001;F =11.192,P ＜ 0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).Under inverted microscope,the cells of the platinum groups were shrunken and round,the volume and quantity were reduced,the morphology was irregular,the gap was increased,and the changes were more obvious with the increase of the concentration and the culture time.The numbers of penetrating cells of the blank group,2,6,12 μg/ml lobaplatin groups were 87.27 ±9.38,71.02 ± 8.92,53.20 ± 10.02 and 21.02 ± 7.37 respectively.The difference was statistically significant (F =87.291,P ＜ 0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).The A values of ERK protein in the blank group,2,6,12 μ~ml lobaplatin groups (0.955 ± 0.021、0.953 ± 0.023、0.950 ± 0.020、0.951 ±0.022)showed no significant difference (F =2.033,P =0.783),but the A values of p-ERK protein in the four groups were 0.941 ±0.015,0.831 ±0.020,0.620 ±0.019 and 0.493 ±0.017 respectively,which showed significant difference (F =11.921,P ＜0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).Conclusion Lobaplatin can inhibit the proliferation and invasion of cervical cancer CaSki cells,which may be related to the inhibition of the expression of p-ERK protein.

Targeted antitumor effects of C-PC/CMC-CD55sp nanospheres on cervical carcinoma Caski cells / 中国肿瘤生物治疗杂志

@# Objective: To prepare a new type of phycocyanin/carboxymethyl chitosan-CD55 ligand peptide (CPC/CMC-CD55sp) nanospheres, and to study its targeted therapeutic effect on cervical cancer Caski cells. Methods: The novel CPC/CMC-CD55sp nanospheres (CPC/CMC-CD55sp) were synthesized by ionic cross-linking method, and the properties of nanospheres were observed by transmission electron microscopy (DLS) and fourier transform infrared spectroscopy (FTIR). The expression of CD55 on the surface of Caski and fibroblast (L-929) cells was detected by Western blotting and flow cytometry. The effect of nanospheres on the proliferation of Caski cells was detected by CCK-8. Flow cytometry and fluorescence microscopy were used to detect the uptake of microspheres by Caski cells; Western blotting and flow cytometry were used to detect the effect of CPC/CMC-CD55sp on expressions of apoptosis-related proteins and apoptosis rate in Caski cells; the hemolysis test was used to determine the biological safety of the drug. Results: CPC/ CMC-CD55sp was successfully prepared with good morphology and uniform diameter; and CD55 was highly expressed on the surface of Caski cells but low expressed on the surface of L-929 cells (P<0.01). CPC/CMC-CD55sp could targeted and efficiently reach Caski cells and be ingested into the cells. It exhibited weak hemolysis effect on human peripheral blood, which was in the safe range. CPC/ CMC-CD55sp displayed obvious inhibitory effect on Caski cell proliferation, and could induce cell apoptosis (P<0.05 or P<0.01). Conclusion: The new CPC/CMC-CD55sp can targeted inhibit the growth of cervical cancer Caski cells via inducing its apoptosis and has good bio-safety, which provides a new idea for the research and development of anti-tumor marine drugs.

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.

Immunogenic apoptosis of human Caski cervical cancer cells induced by Lidamy-cin / 中国免疫学杂志

Objective:To investigate effects of Lidamycin (LDM,C-1027) on the proliferation and immunogenic transform of human Caski cervical cancer cells and to provide the basic experiment data and theoretical supports for establishment of the new immu-notherapy method mediated by LDM. Methods:MTT was used for the analysis of cell proliferation;apoptosis rate was analyzed by flow cytometry;Western blot was used to analyze the effect of LDM on the expression of Bax and Bcl-2 in Caski cells;the Flow cytometry was used to detect the expression of Calreticulin ( CRT ) on the cell surface. Results: Lidamycin inhibited proliferation of Caski cells significantly in the time-and dose-dependent manners;The apoptotic cell ratio induced by 5 μg/L Lidamycin was 11. 5% ,Comparing with the control group, Lidamycin treatment increased Bax but decreased Bcl-2 contents significantly within Caski cells, it also significantly increased the expression of CRT on the cell surface of Caski cells from 2. 31% to 67. 2%. Conclusion: Lidamycin has pharmacological activity in inhibiting proliferation of the human cervical Caski cells and the underlying mechanism is related with inducing the intrinsic mitochondrial pathway of apoptosis. In the same time,Lidamycin can increase the expression of CRT on the cell surface,so it may have the ability to promote the immunogenic apoptosis of tumor cells.

Construction of expression vector containing rat rCB1 gene and its influence on the apoptosis in human cervical cancer CaSki cell line / 中国实验动物学报

Objective To construct rCB1 gene eukaryotic expression vector, detect its expression in the cell, and explore its influence on apoptosis in human cervical cancer CaSki cells.Methods The total RNA was extracted from rat brains.The rCB1gene was amplified by RT-PCR.The pcDNA3.1(+)-rCB1 was constructed by enzyme digestion, purifi-cation, bind the PCR purification products and pcDNA3.1 (+) DNA.The pcDNA3.1 (+)-rCB1 plasmid was transfect-ed into HEK293 and CaSki cells by liposomes.The expression and localization of rCB1 were detected by Western blot and immunofluorescence combined with confocal laser scanning microscopy.The apoptosis rate of CaSki cells was detected by flow cytometry.The expression of rCB1, Bcl-2, Bax and Bad was detected by Western blot and real-time fluorescence quantitative RT-PCR (qRT-PCR).Results The 5300 bp pcDNA3.1(+) and 1500 bp rCB1 were obtained after diges-ting the pcDNA3.1 ( +)-rCB1.The result of sequencing was in agreement with the sequence of rCB1 gene ( NM_012784.4 ) .The rCB1 expressed in the membrane and cytoplasm when pcDNA3.1 (+)-rCB1 plasmid was transfected into HEK293 cells.The apoptosis rate of rCB1 group was increased compared with the blank group when pcDNA3.1 (+)-rCB1 plasmid was transfected into CaSki cells (P<0.05).Compared with the blank group, rCB1 gene upregulated the expres-sion of Bax and Bad, and suppressed the expression of Bcl-2.The statistical difference was significant ( P <0.05). Conclusions The pCDNA3.1(+)-rCB1 eukaryotic expression vector is constructed successfully.It is found that rCB1 is expressed in membrane and cytoplasm of HEK293 cells.rCB1 can significantly promote the apoptosis in cervical cancer CaSki cells by up-regulating the expression of Bax and Bad, and down-regulating the expression of Bcl-2 as well.

Induction of cytotoxic T lymphocytes against cervical cancer cell line CaSki using therapeutic dendritic cell vaccine / 肿瘤

Objective: To observe the inductive effect of cytotoxic T lymphocytes (CTLs) against human cervical cancer cell line CaSki using therapeutic dendritic cells (DCs) vaccine in vitro . Methods:. Immature mouse DCs were isolated and cultured. The expressions of cell-surface CD40, CD86, major histocompatibility complex (MHC)-. and CD11c in immature DCs were detected by flow cytometry (FCM). Then the immature mouse DCs were infected with recombinant adenoviral vector carrying human papillomavirus (HPV )16 E 6/E 7 (pAd-E6/E7), and the CaSki cell lysate-loaded autologous DCs vaccine was prepared. The expression of green fluorescent protein in pAd-E6/E7-infected immature mouse DCs was observed under a laser scanning confocal microscope, and the expression of E6 protein was detected by Western blotting. DCs vaccine was used to induce specific CTLs, were subsequently co-cultured with CaSki cells. The killing effect of CTLs against CaSki cells was determined using cell counting kit8(CCK8) assay. Results: HPV16 E6/E7-specific DCs vaccine was successfully prepared. CTLs which induced by DCs vaccine exerted a killing effect on CaSki cells. This killing effect was higher in pAd-E6/E7-infected group than those in CaSki cell lysate-loaded group and the untreated control group (P<0.05). Conclusion: Genetically modified DC vaccine can successfully be prepared by infection with pAd-E6/E7, and it has a significant effect on triggering of specific CTLs against CaSki cells.

The mechanism of cell death in etoposide treated cervical cancer cells: apoptotic and non-apoptotic programmed cell death / 부인종양

OBJECTIVE: Etoposide is a potent and widely used antineoplastic agent. It is able to induce apoptosis in most cell types. However, very little is known about its mechanism of action. In this study, we demonstrate the cytotoxic signal that induced by etoposide and investigate how etoposide exerts antitumor activity in HPV-16 (+) CaSki cervical carcinoma cells. METHODS: Antiproliferation activity was measured in CaSki cell lines by using MTT assays, DNA fragmentation assay. Cell cycle distribution was analyzed using flow cytometry. Expression of proteins involved in the apoptotic pathway was analyzed by Western blotting (WB). Electron microscopic (EM) and biochemical studies (Western blotting, RT-PCR) revealed that non-apoptotic death was associated with autophagosomes/-autolysosomes. These parameters have also been measured in cells treated with 3-methyladenine (an autophagy inhibitor), zVAD-fmk (a pan-caspase inhibitor) and both. RESULTS: The etoposide induced apoptosis. In cell cycle analysis, etoposide-treated CaSki cells were few induced hypodiploid DNA content, suggesting that apoptotic cell death. EM study revealed that autophagic appearance in the presence of etoposide exhibited by autophagosomes/autolysosomes. It was confirmed by LysoTracker probe and WB against Beclin 1, APG 5, APG 12 and p53. When autophagy was blocked by 3-MA, not only the protein expression of Beclin 1, but also the antitumor effect of etoposide was suppressed. On the other hand, the addition of zVAD-fmk could induce a few etoposide-induced autophagy. And etoposide-treated CaSki cells were rescued by combination of 3-MA and zVAD-fmk. CONCLUSION: Our results suggest that etoposide not only initiated apoptosis but ultimately caused cell death through autophagy. In this study, we demonstrate novel features for the action of etoposide in HPV-16 (+) CaSki cervical carcinoma cells. Autophagic cell death induction by some anticancer agents underlines the potential utility of its induction as a new cancer treatment modality.

Effect of S100A8/A9 Protein Complex on F-actin Network in Human Cervical Carcinoma Cell Line,CasKi Cells / 中国微创外科杂志

Objective To investigate the effect of S100A8/A9 protein complex on the surface morphology and the F-actin network in human cervical carcinoma cell line,CasKi cells.Methods After being cultured with 20 ?g/ml S100A8/A9 protein complex,the cell skeleton of the CasKi cells were observed under a confocal scanning fluorescence microscope by staining the F-actin network.Atomic force microscopy(AFM) was employed to reveal the change of ultrastructure of the cell surface in vivo.ResultsAfter being cultured with the S100A8/A9 protein complex for 24 hours,the F-actin network disorder was revealed.Most of the F-actins distributed peripherally.The OD value of the F-actin decreased significantly from 92.42?5.16 to 57.67?3.70 after been treated with the S100A8/A9(t=5.268,P=0.000).The AFM showed a withdrawing morphology with reduced pseudopodia and destruction of stress fibers. Conclusion S100A8/A9 protein complex can change the ultrastructure of the surface of CasKi cells and its stress fibers by re-distributing of the F-actin in the cells.

Vaccine preparation of dendritic cell transfected with HPV16E6 antigen gene and its biological characteristics / 中国免疫学杂志

Objective:To prepare the vaccine of DC derived from human peripheral blood and transfected with HPV16E6 antigen gene, and to detect its morphological character,surface marker and immunological effect.Methods:DC-enriched populations were prepared from human peripheral blood mononuclear cell(PBMC) with the combination of rhGM-CSF,rhIL-4 and rhTNF-?. The plasmid containing HPV16E6 gene was transfected into DC with lipofectamine. The morphology of DC was observed dynamically, and the expression of surface markers of DC vaccine could be detected using immuno-cytochemical staining and flow cytometry. MTT assay was applied to detect the activity of CTL in vitro.Results:The transfected DC had typical morphologic and phenotypic characteristics, and expressed E6 protein 47.3%, CD80 82.5%, CD86 79.8% and CD83 85.7%. The killing activities of CTL to Caski cells induced by transfected DC were higher evidently than that of control groups(P

Combined Effect of Radiation and Tyrphostin AG 1478 in Human Cervical Cancer Cell Lines / 대한산부인과학회잡지

OBJECTIVE: We studied the possibility that addition of Tyrphostin AG 1478 which is selective epidermal growth factor inhibitor, would enhance the effect of radiation on human cervical cancer cell lines, HeLa and CaSki. METHODS: Tyrphostin was added to the cells which were irradiated. The ratio of dead cells was estimated by trypan blue dye examination, and survived cell fractions were estimated by clonogenic assays. The presence and degree of apoptosis were examined by DNA electrophoresis and nuclear dye using propium iodide. RESULTS: The growth was completely inhibited in both cell lines, but the addition of tyrphostin resulted in different effects on the radiation induced cell death and apoptosis in each cell line. However, the percentage of dead cells and apoptotic cells was decreased in HeLa cell line compared with CaSki cell line. The ultimate survived cell fractions determined by clonogenic assays were decreased in both cell lines and the size of colony was also decreased. CONCLUSION: These data suggest that the addition of Tyrphostin is able to increase the radiotherapeutic effects on human cervical cancer cells, and this synergistic effect may result from effective blocking of radiation-induced accelerated repopulation of cancer cells by tyrphostin.

Expression of c-jun by X-ray According to Cell Growth State in CaSki Cell Line / 대한방사선종양학회지

PURPOSE: The expression pattern of c-jun by ionizing radiation according to cell growth state (exponential growth phase vs. stationary phase) and its relationship with cell cycle redistribution were investigated. MATERIALS AND METHODS: The exponential growth phase (day 4) and stationary phase (day 9) cells were determined from cell growth curve according to the elapse of days in CaSki. The cells were irradiated using 6 MV X-ray with a dose of 2 Gy at a fixed dose rate of 3 Gy/min. Northern blot analysis was performed with total cellular RNA and cell cycle distribution was analyzed using flow cytometry according to time-course after irradiation. RESULTS: The maximum expression of c-jun occurred 1 hour after irradiation in both exponential growth and stationary phase cells. After then c-jun expression was elevated upto 6 hours in exponential growth phase cells, but the level decreased in stationary phase cells. Movements of cells from G0-G1 to S, G2-M phase after irradiation were higher in exponential growth phase than stationary phase. CONCLUSION: c-jun may be involved in the regulation of cellular proliferation according to the growth states after irradiation.