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Objective Study on the effect of isocorydine in human cervical cancer Siha cells xenografts in nude mice , to explore the inhibition mechanism of isocorydine in cervical carcinoma .Methods Establishment of human cervical cancer Siha cells subcutaneous xenografts model in BALB /c (nu/nu) nude mice.When the average diameter of the transplanted tumor≥0.5 cm, mice were randomly assigned into control group and experimental group .In experimental group, the model was administered by intraperitoneal injection of different doses of isocorydaline .After 4 weeks, the tumor tissues were removed , the histopathological changes of the tumor were observed by HE staining , and the ex-pression of proteins in the tumor tissue were observed by immunohistochemical staining and Western blot .Results Compared with the control group , the tumor volume of experimental group was significantly decreased ( P<0.05);the cell morphology transformed from stem cells to epithelioid cells , and the expression of E-cadherin was significantly in-creased while the expression of HPV 16E6 and vimentin was decreased .Conclusions Isocorydine may inhibit the de-velopment of cervical cancer by inhibiting the expression of E 6 protein and EMT-related signaling pathway .
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OBJECTIVE To explore the pro-apoptotic mechanism of juglone in SiHa cells and to in?vestigate its associated signal transduction pathways. METHODS SiHa cells were treated with juglone 20μmol·L-1 for 24,48 and 72 h. Cellular morphology was detected by inverted microscopy.The cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay. After 24 h treatment with juglone 20μmol·L-1,the cell apoptosis was detected by flow cytometry while the expressions of apopto?sis-related protein BCL-2,BAX and cleave-caspase-3,PI3K/AKt pathway-related protein PTEN,AKT and pAKT were detected by Western blotting. RESULTS After treatment with juglone for 24, 48 and 72 h,the growth of SiHa cells was significantly inhibited. Compared with cell control group,cells in juglone treated gruop were sparse,slipped off the wall,became round and the cell proliferation inhibitory rate was 43.3%,63.0%and 73.1%(P<0.05,P<0.01),respectively. Twenty-four hours post treatment, the early apoptosis rate of juglone treated gruop cells was increased by(6.47±1.79)%(P<0.01)compared with cell control group. Western blotting results showed that the expression of BCL-2 decreased by 53.0%while the expression of BAX and caspase-3 increased by 85.5%and 183.3%,respectively. The expression of PTEN was increased by 75.0% but the pAKt was decreased by 45.8%(P<0.01). CONCLUSION Juglone can upregulate the expression of PTEN, thus inhibiting PI3K/AKt pathway and promoting apoptosis of SiHa cells.
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AIM:To explore the effect of peptidyl-prolyl cis/trans isomerase (Pin1) inhibitor juglone on apop-tosis of human cervical cancer SiHa cells.METHODS:Cultured SiHa cells were incubated with juglone at concentrations of 10, 20, 50, 80 and 100 μmol/L for 24 h.The SiHa cell activity was detected by methyl thiazolyl tetrazolium ( MTT) assay.The cell apoptosis was analyzed by flow cytometry with Hoechst 33258 staining.The protein levels of cleaved caspase-3,8,9 and PTEN was determined by Western blotting.RESULTS:In different doses of juglone groups, the SiHa cell growth was greatly inhibited ( P<0.05) in a dose-dependent manner as compared with control group.The IC50 of ju-glone was 20.4 μmol/L.After treatment with juglone at concentration of 20 μmol/L for 12 h, the apoptosis of SiHa cells was induced, and the typical morphological changes of cell apoptosis such as karyopyknotic pyknic hyperfluorescence bolus, nuclear fragmentation and apoptotic body were observed by Hoechst 33258 staining.The early apoptotic rate was increased significantly as compared with the control.The protein levels of cleaved caspase-3, 8, 9 and PTEN were also increased sig-nificantly as compared with control group.CONCLUSION:Juglone significantly inhibits the cell activity and induces the apoptosis of SiHa cells in vitro by inhibiting the caspase pathway and increasing the expression of anti-oncogene.
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Objective To study the antitumor effect of new photodynamic therapy (PDT) applying TMPyP4 combined with nucleolin silence on cervical cancer SiHa cells in vitro, and to explore an available combination treatment project for cervical cancer.Methods The SiHa Cells were divided into blank control group, RNAi group,PDT group and PDT-RNAi group.The proliferation activities of SiHa cells were assessed by Cell Counting Kit-8 (CCK8)assay.The apoptotic rates were measured by flow cytometry (FCM)with Annexin Ⅴ/PI staining. The invasiveness abilities were assessed by Transwell assay. Results Compared with blank control group, the inhibitory rates of growth of SiHa cells in RNAi group, PDT group and PDT-RNAi group were increased significantly (P<0.01);the inhibitory rates of growth of SiHa cells in PDT-RNAi group were higher than those in PDT group and RNAi group (P<0.05 ). The Q value was 1.27. Compared with blank control group, the apoptotic rates of SiHa cells in experiment groups were increased (P<0.05 ), and the late apoptotic rate in PDT-RNAi group was also increased;there were significant differences of the apoptotic rates between PDT-RNAi group and RNAi group, PDT group (P<0.05 ). Compared with blank control group, the cell invasiveness abilities of SiHa cells in experiment groups were decreased;there were significant differences of the invasiveness abilities of SiHa cells between PDT group and RNAi group (P<0.05).Conclusion New PDT shows a strong photodynamic effect on the SiHa cells,which can inhibit the proliferation and invasiveness and induce the apoptosis of SiHa cells invitro;nucleolin silence shows a good synergy effect to PDT.
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Objective To observe the changes of cell cycle on cancer cells after dihydroartemisinin and X-ray irradiation. Methods Human HeLa cells of cervical cancer with p53 mutation was used and human SiHa cells of cervical cancer with wild p53 was used as control. Flow cytometry was used to detect the effect of dihydroartemisinin (20 and 100 μmol/L) and irradiation (6 Gy)on cell cycle. Western blot was used to measure the levels of cell cycle protein. Results G2 arrest was observed in irradiated HeLa cells, which the proportion of cells in G2 phase was increased from 14.45% to 73. 58% after 6 Gy X-ray irradiation, but it was abrogated by dihydroartemisinin from 73. 58% to 48.31%in HeLa cells, and it had no change on the SiHa cells. The elevated Weel protein and the lowered Cyclin B1 protein were observed with the G2 arrest severity. The expression of radiation-induced Weel protein was suppressed and the Cyclin B1 protein was increased after dihydroartemisinin treatment, which was in accordance with the abrogation of radiation-induced G2 delay. Conclusions The main effect of irradiation on cell cycle of p53 mutated HeLa cells is G2 arrest. Dihydroartemisinin could abrogate it, which is associated with the changes of Weel protein and Cyclin B1 protein. In Siha cells, the main effect of irradiation on cell cycle is G1 arrest, and dihydroartemisinin has no effect on it.
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Functional defects in mitochondria are involved in the induction of cell death in cancer cells. We assessed the toxic effect of camptothecin against the human cervical and uterine tumor cell line SiHa with respect to the mitochondria-mediated cell death process, and examined the combined effect of camptothecin and anticancer drugs. Camptothecin caused apoptosis in SiHa cells by inducing mitochondrial membrane permeability changes that lead to the loss of mitochondrial membrane potential, decreased Bcl-2 levels, cytochrome c release, caspase-3 activation, formation of reactive oxygen species and depletion of GSH. Combination of camptothecin with other anticancer drugs (carboplatin, paclitaxel, doxorubicin and mitomycin c) or signaling inhibitors (farnesyltransferase inhibitor and ERK inhibitor) did not enhance the camptothecin-induced cell death and caspase-3 activation. These results suggest that camptothecin may cause cell death in SiHa cells by inducing changes in mitochondrial membrane permeability, which leads to cytochrome c release and activation of caspase-3. This effect is also associated with increased formation of reactive oxygen species and depletion of GSH. Combination with other anticancer drugs (or signaling inhibitors) does not appear to increase the anti-tumor effect of camptothecin against SiHa cells, but rather may reduce it. Combination of camptothecin with other anticancer drugs does not seem to provide a benefit in the treatment of cervical and uterine cancer compared with camptothecin monotherapy.
Subject(s)
Humans , Apoptosis , Camptothecin , Carcinoma, Squamous Cell , Caspase 3 , Cell Death , Cell Line , Cell Line, Tumor , Cytochromes c , Doxorubicin , Membrane Potential, Mitochondrial , Mitochondria , Mitochondrial Membranes , Mitomycin , Paclitaxel , Permeability , Reactive Oxygen Species , Uterine NeoplasmsABSTRACT
The effects of nanometer realgar suspension on proliferation and apoptosis of human uterine cervix cancer cell line SiHa cells and oncogenic genes HPV16E6/E7 were investigated. A "micro-jet efflux" strategy was used for the preparation of nanometer realgar suspension. SiHa cells were treated with nanometer Realgar suspension in various concentrations (6.25,12.5,25 and 50mg/L) for different durations (12,24,48 and 72h). The inhibitive effect of nanometer realgar suspension on growth of SiHa cells was detected by MIT method. Special morphological changes of apoptosis were observed by transmission electron microscopy (TEM) and DNA fragments electrophoresis. The apoptotic rate was quantified by flow cytometry (FCM). The expression of HPV16E6/E7 mRNA and protein was assayed by RT-PCR and Western blot respectively. The results showed after being treated with 25-50mg/L nanometer realgar suspension for 48h, the survival rate of SiHa cells was decreased, and apoptotic rate markedly increased in a time- and concentration-dependent manner. TEM and DNA electrophoresis revealed the special morphological changes of apoptosis. The apoptotic rate of SiHa cells treated with nanometer realgar suspension was significantly higher than in the control group (P<0.01), and G0/G1 phase arrest appeared following treatment with nanometer realgar suspension in 25 and 50mg/L for 48h.RT-PCR and Western blot assay indicated that nanometer realgar suspension reduced the HPV16E6/E7 gene expression. Nanometer realgar suspension could inhibit the proliferation and induce apoptosis of SiHa cells. The mechanism may be related to the down-regulation of the HPV16E6/E7 gene expression.
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Objective To discuss the effect of Qingdushuan on SiHa cells of cervical cancer proliferation and protein expression in PTEN-MDM2-p53 network from the level of cell biology and molecular biology. Methods SiHa cells were cultivated with 4% containing serum. Normal control group, blank serum group, Qingdushuan group, Baofukang group and interferon group were established. The influence of inhibiting and proliferating SiHa cell was detected by MTT assay. Three protein expression was detected by Western blot method. Results After cultured with drug containing serum, the number of cells decreased, 4% concentration of serum for 72 h was the strongest. Compared with the control group, there were significant differences (P