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.

Effects of ampelopsin on proliferation, cell cycle and apoptosis of human cervical cancer SiHa cells / 中国药理学通报

Aim To investigate the effects of ampelopsin (AMP) on proliferation, cell cycle and apoptosis of human cervical cancer SiHa cells, and its possible mechanism of action. Methods MTT assay was used to detect the inhibitory effect of AMP with different concentrations (10, 20, 40, 80, 160, 320 μmol • L

Ellagic acid promotes apoptosis of human cervical cancer cells through endoplasmic reticulum stress and mitochondrial damage / 中国药理学通报

Aim To investigate the effect of ellagic acid on human cervical cancer cells and its correlation with endoplasmic reticulum stress ( ERS) - mitochondrial apoptosis pathway. Methods Hela cells were treated with ellagic acid, and cell survival rate, apoptosis and expression of ERS mitochondrial apoptosis pathway protein were detected. After pretreatment with ERS inhibitor 4-PBA, the expression of mitochondrial apoptosis pathway protein was detected. Results Ellagic acid showed concentration-dependent and time- dependent killing effect on Hela cells, and could significantly increase the apoptosis of Hela cells. Ellagic acid significantly increased the expression levels of ERS related proteins (GRP78, CHOP, p-PERK, p-IREl) in Hela cells, and significantly increased the expression levels of mitochondrial proapoptotic proteins . Bax and cleaved caspase-3, while significantly decreased the expression level of mitochondrial anti apop- totic protein Bcl-2. Another group of experiments showed that pretreatment with ERS inhibitor 4-PBA could significantly reduce the expression levels of Bax and cleaved caspase-3, and significantly increase the expression level of Bcl-2, which was close to the normal group. Conclusions Ellagic acid can significantly promote the apoptosis of human cervical cancer cells, and its mechanism is related to ERS mitochondrial apoptosis pathway.

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.

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.

Establishment of Paclitaxel-resistance Cervical Carcinoma Cell Line Hela/PTX and Study on its Biological Characteristics / 中国药师

Objective:To establish a paclitaxel (PTX)-resistance cell line of human cervical carcinoma Hela/PTX and investigate its biological features. Methods:Using Hela as the negative cells,drug-resistant cervical cancer Hela/PTX cells were applied as the target cells. The morphology changes were observed under an optical microscope;the drug resistance of Hela/PTX to different chemotherapeutic drugs was detected by CCK-8 method. The accumulation of drugs in the cells was determined by HPLC. The expression of P-glycoprotein (P-gp), Survivin protein and mRNA was detected by Western blot and RT-PCR. Results:Hela/PTX multidrug resistant cell line was successfully established. The resistance index to PTX was 58.40 and the resistance index of cross-resistance to docetaxel and 5-fluorouracil was 8.11 and 4.62,respectively. The mRNA expression level of P-gp and Survivin protein in Hela/PTX cells respectively was(9.93 +0.90) times(P < 0.05) and(3.02 +0.57) times(P <0.05) of that in Hela cells. The expression of P-gp and Survivin protein in drug-resistant cells was significantly higher than that in parental cells.Conclusion:Human cervical cancer resistant cell line Hela/PTX with the basic biological characteristics of multidrug resistance is successfully established, which can be used as an in vitro model for the studies on paclitaxel resistance mechanism and provide experimental basis for the studies of multidrug resistance mechanism of cervical cancer.

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.

Study on Effects of Podophyllotoxin Derivative QW-83 on Human Cervical Cancer HeLa Cell Apoptosis and Its Mechanism / 中国药房

OBJECTIVE:To study the effects of podophyllotoxin derivative QW-83 on human cervical cancer HeLa cell apopto-sis and its mechanism. METHODS:After treated with 0(negative control),0.01,0.1,1 and 10 μmol/L QW-83 and positive drug etoposide(VP-16)for 48 h,proliferation inhibition rate and IC50 of HeLa cell were determined by MTT assay. The morphological changes of HeLa cell were observed by Hochest 33342 staining after treated with QW-83 [0(negative control),2.5,5,10μmol/L] for 48 h;flow cytometry was used to detect apoptosis rate;semi quantitative RT-PCR was adopted to detect the expression of apop-tosis related gene P53,Bax,Casepase-3,Casepase-8,Casepase-9 and Bcl-2 mRNA. RESULTS:Compared with negative control, 1,10 μmol/L VP-16 and QW-83 had obvious proliferation inhibition effect on HeLa cells (P<0.05 or P<0.01),and IC50 were (5.11±0.43)μmol/L and(4.96±0.54)μmol/L. Hochest 33342 staining results showed QW-83 could obviously induce cells apopto-sis and nuclear pyknosis. Flow cytometry showed QW-83 could increase apoptosis rate in concentration-dependent manner,being 16.89%-62.56%. RT-PCR showed mRNA expression of P53,Bax,Caspase-3,Casepase-8 and Casepase-9,Bcl-2/Bax increased, while mRNA expression of Bcl-2 decreased after treated with QW-83(P<0.05). CONCLUSIONS:Podophyllotoxin derivative QW-83 can induce HeLa cell apoptosis,and its mechanism may be associated with regulate mRNA expression of apoptosis related gene.

Study on the Mechanism of Autophagy of HeLa Cells in Human Cervical Cancer Induced by Fucoxanthin / 中国药房

OBJECTIVE:To study the mechanism of autophagy of HeLa cell in human cervical cancer induced by fucoxanthin. METHODS:MTT was adopted to determine the cell activity and calculate the inhibition rate after HeLa cells were cultured by 1, 10,20,40 and 80 μmol/L of fucoxanthin for 48 h. Flow cytometry was used to determine the cell cycle and apoptosis rate after HeLa cells were cultured by 0(blank control),10,20 and 40 μmol/L of fucoxanthin for 48 h;acridine orange staining,LysoTrack-er Red staining,HeLa-GFP-LC3 method and fluorescence microscope were used to observe the autophagy state;Western blot was used to determine the expressions of proteins related to autophagy. RESULTS:The cells had obvious inhibition effect on the cell growth after being cultured by 0,10,20,40 and 80 μmol/L of fucoxanthin. The cell was blocked in G0/G1 stage after being cul-tured by 10,20 and 40μmol/L of fucoxanthin,and had no obvious effect on the apoptosis rate;autophagy degree was increased af-ter the cells were cultured by 40 μmol/L fucoxanthin for 48 h. Compared with blank control,40 μmol/L fucoxanthin could promote LC3Ⅰ transferring into LC3Ⅱ and the expressions of Beclin-1,PTEN,p21;and inhibit the phosphorylation of p-Akt,p-p70S6K and p-mTOR. The pre-treatment by autophagy inhibitor 3-methyladenine(5 mmol/L)could reverse the autophagy of HeLa cells in-duced by fucoxanthin;U0126 could partly reverse the autophagy of HeLa cells induced by fucoxanthin. CONCLUSIONS:Fucoxan-thin can induce the authphagy of HeLa cells by inhibiting Akt signaling pathway and activating MEK/ERK signaling pathway.

Study effect of shikonin on proliferation of human cervical cancer Hela cells and its mechanism / 中国生化药物杂志

Objective To study effect of Shikonin on human cervical cancer Hela cell growth suppression in vitro and its mechanism. Methods MTT assay was used to examine the growth inhibition of Shikonin in Hela cells.And then, the measurement of both ROS Levels and Mitochondrial Membrane Potential (ΔΨm ) were performed to clarify the mechanism of antitumor in Hela cells by Shikonin.Results Shikonin significantly inhibited the growth of Hela cells in a concentration-dependent manner.Shikonin increased generation of en-dogenous reactive oxygen species ( ROS) and decreased mitochondrial membrane potential.Furthermore, anti-oxidants N-acetylcysteine ( NAC) could significantly reduce the antitumor activity of SK in Hela cells.Conclusion These results suggest that mitochondrial aerobic respiration shift and endogenous ROS augmentation contribute to the action of Shikonin against Hela cells.

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.

Cytotoxic activity of kenaf (Hibiscus cannabinus L.) seed extract and oil against human cancer cell lines

Objective: To examine the cytotoxic properties of both the kenaf (Hibiscus cannabinus L.) seed extract and kenaf seed oil on human cervical cancer, human breast cancer, human colon cancer and human lung cancer cell lines.Methods:kenaf seed oil on human cancer cell lines was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and sulforhodamine B assays. Cell morphological changes were observed by using an inverted light microscope.Results:The in vitro cytotoxic activity of the kenaf (Hibiscus cannabinus L.) seed extract and cancer cell lines. Morphological alterations in the cell lines after KSE and KSO treatment were observed. KSE and KSO possessed effective cytotoxic activities against all the cell lines been selected.Conclusions:KSE and KSO could be potential sources of natural anti-cancer agents. Further The kenaf seed extract (KSE) exhibited a lower IC50 than kenaf seed oil (KSO) in all of the investigations on using kenaf seeds for anti-proliferative properties are warranted.

Cytotoxic activity of kenaf (Hibiscus cannabinus L.) seed extract and oil against human cancer cell lines

<p><b>OBJECTIVE</b>To examine the cytotoxic properties of both the kenaf (Hibiscus cannabinus L.) seed extract and kenaf seed oil on human cervical cancer, human breast cancer, human colon cancer and human lung cancer cell lines.</p><p><b>METHODS</b>The in vitro cytotoxic activity of the kenaf (Hibiscus cannabinus L.) seed extract and kenaf seed oil on human cancer cell lines was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and sulforhodamine B assays. Cell morphological changes were observed by using an inverted light microscope.</p><p><b>RESULTS</b>The kenaf seed extract (KSE) exhibited a lower IC50 than kenaf seed oil (KSO) in all of the cancer cell lines. Morphological alterations in the cell lines after KSE and KSO treatment were observed. KSE and KSO possessed effective cytotoxic activities against all the cell lines been selected.</p><p><b>CONCLUSIONS</b>KSE and KSO could be potential sources of natural anti-cancer agents. Further investigations on using kenaf seeds for anti-proliferative properties are warranted.</p>

Study on methylation status of RASSFIA gene promoter and exon 1 in cervical cancer cell lines / 中国癌症杂志

Background and purpose:Loss or altered expression of Ras association domain family 1A gene (RASSF1A) through DNA methylation has been associated with the pathogenesis of a variety of cancers, which suggests the tumor suppressor function of this gene. This study aimed to explore the effect of DNA methyltransferase inhibitor 5-Aza-2’deoxycytidine (5-Aza-dc) on demethylation and expression of RASSF1A in cervical cancer cell lines. Methods:HPV positive cervical cancer cell lines HeLa and Caski, HPV negative cell lines HT-3 and C-33A were treated with two different concentration of 5-Aza-dc (5 μmol/L, 10 μmol/L). MSP (methylation-specific PCR) and Bisulfite genomic sequencing PCR (BGS) combined with TA clone were used to investigate methylation status of RASSF1A gene promoter and exon 1 before and after treatment of 5-Aza-dc. RASSF1A gene mRNA expression was detected by RT-PCR. Results:Two HPV positive cell lines showed hypomethylated RASSF1A promoters and expressed RASSF1A mRNA, and after treatment with 5-Aza-dc, the mRNA expression of RASSF1A did not change significantly (FHeLa=3.003, P=0.125; FCaski=0.045, P=0.956). Two HPV negative cervical cancer cell lines showed hypermethylation status of RASSF1A promoter and silenced RASSF1A. After treatment with 5-Aza-dc, demethylation occurred in the promoter region of RASSF1A gene, which subsequently induced re-expression of this gene in HT-3 and C-33A. The F test (FHT-3=18.002, P=0.03;FC-33A=17.179, P=0.03) and LSD-t test (P<0.05) demonstrated that significant difference in the expression of RASSF1A was found upon two different concentrations drug treatment.Conclusion:The methylation status of promoter and exon 1 of RASSFIA gene in HPV positive and HPV negative cervical cancer cell lines are different. The promoter hypermethylation is correlated with RASSF1A gene expression in HPV negative cervical cancer cell line HT-3 and C-33A, and plays a key role in RASSF1A silencing. 5-Aza-dc may effectively reverse the methylation status of RASSFIA gene promoter in cervical cancer HT-3 and C-33A cells and reactivate gene expression silenced by aberrant hypermethylation in a dose-dependent manner within certain extent.

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.

Human cervical cancer oncogene and tumor / 国际肿瘤学杂志

Human cervical cancer oncogene(HCCR) is newly identified in cervical cancer tissues,and expresses in most human tumors. Resarches show that HCCR is a candidate marker for human hepatocelular carcinoma and breast cancer. Moreover,the expression of HCCR is regulated by mitogen-activated protein kinase (MAPK)and phosphatidylinositol 3-kinase signal pathway. The proliferation,apoptosis,migration and invasion of tumor cells could be inhibited by siRNA of HCCR.

Induction of apoptosis in oral cancer cell line through an recombinant HCCS-1 adenovirus

Despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with oral squamous cell carcinoma has not significantly improved over the past several decades. Gene therapy is currently under investigation and shows us new possibility of cancer curing method. This experiment was undergone to find out the cell growth inhibition effect and evidence of apoptosis by HCCS-1(human cervical cancer suppressor-1), one of the candidates of tumor suppressor gene, transducted to human oral cancer cell line. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transducted with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the transfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay using cell count kit(CCK). To show the evidence of apoptosis, DNA fragmentation assay and flow cytometry(FACS) were performed. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1), and importation efficiency was 20% at 2 MOI(multiplicity of infection), 80% at 20 MOI. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transducted cell lines. As a result of CCK, when comparing to control subjects, transducted group showed 50% growth inhibition. In DNA fragmentation assay, according to increasing of MOI, DNA volume was diminished. In FACS analysis, DNA distribution showed fragmentation.This results imply that HCCS-1gene has growth inhibition effect in human oral cancer cell lines through apoptosis induction.

The experimental investigation derived from Sdosodine Hydrochlorida on apoptosis of the cervical cancer ME180 cells / 中国免疫学杂志

Objective:To experimental study the inhibiting effects of SBHL on the growth of cervical cancer cells and to investigate the mechanisms.Methods:The proliferation of ME180 cells was observed by MTT assasy; The changes of cells cycles and apoptosis of ME180 cells treated with SBHL were analyzed by agar gel electrophoresis and FCM assay;. The changes of the ultra structure of cells was observed by Transmission eleconmicrograph.Results:The proliferation of ME180 cells was inhibited after treating on SBHL for 24 h.The inhibiting effect increased granually following the concentration of SBHL(P