Objective: To discuss the the effect of FGFR4 gene silencing on growth, metastasis and the cis-platinum (DDP) chemotherapy sensitivity of Hela cervical cancer cells. Methods: The Hela cervical cancer cells were cultur...Objective: To discuss the the effect of FGFR4 gene silencing on growth, metastasis and the cis-platinum (DDP) chemotherapy sensitivity of Hela cervical cancer cells. Methods: The Hela cervical cancer cells were cultured and divided into 5 groups: blank control group, negative control group, FGFR4-siRNA group, DDP group and FGFR4-siRNA+DDP group. The siRNA targeted to FGFR4 was constructed and transfected into the cells in FGFR4-siRNA group and FGFR4-siRNA+DDP group, and DDP (10 μg/mL) was added into the DDP group and FGFR4-siRNA+DDP group. The growth inhibition ratio was detected by MTT and the apoptosis rate was detected by FCM after 24 h, 48 h and 72 h culturing respectively. The mRNA and protein expression level of FGFR4, VEGF and MMP2 were detected by RT-PCR and western blot respectively. Results: Compared with the blank control, the proliferative activity of FGFR4-siRNA group decreased after the FGFR4 being silenced, and the growth inhibition ratio and apoptosis rate increased, the FGFR4 did almost not express and the expression level of VEGF and MMP2 decreased.The growth inhibition ratio and apoptosis rate of FGFR4-siRNA+DDP group were higher than that of FGFR4-siRNA group and DDP group, the FGFR4 did almost not express either and the the expression level of VEGF and MMP2 were lower than the FGFR4-siRNA group and DDP group. Conclusion: It can be obtained that silencing the FGFR4 of cervical cancer could inhibit cell growth, promote cell apoptosis and increase the chemosensitivity, down-regulate the expression of VEGF and MMP2 to inhibit the ability of invasion and metastasis, which could be one of the targeted therapeutic targets of cervical cancer.展开更多
Objective:To study the inhibitory effect of docetaxel(DOC),oxaliplatin(OXA) and their combination on proliferation of cervical cancer line HeLa.Methods:Cell morphological changes were observed by inverted phase contra...Objective:To study the inhibitory effect of docetaxel(DOC),oxaliplatin(OXA) and their combination on proliferation of cervical cancer line HeLa.Methods:Cell morphological changes were observed by inverted phase contrast microscope,cell inhibition rates in different groups were examined by MTT,and cell cycle and apoptosis rates were determined by flow cytometry(FCM).Results:DOC and OXA inhibited the proliferation of cervical cancer cell line HeLa in a dose-dependent,and combination of the two drugs had an enhanced inhibitory effect;the apoptosis rate was also significantly increased when the two drugs were used in combination.Conclusion:DOC and OXA can synergistically inhibit the proliferation of cervical cancer cell line HeLa,which indicates that combination of the two drugs might has a promising future for clinical treatment of cervical cancer.展开更多
OBJECTIVE To establish a murine uterine cervical cancer cell line and to define its biological characters. METHODS Transplanted tumor tissue was used for in vitro primary culture of U14 cervical carcinoma cells. After...OBJECTIVE To establish a murine uterine cervical cancer cell line and to define its biological characters. METHODS Transplanted tumor tissue was used for in vitro primary culture of U14 cervical carcinoma cells. After 20 passages, we examined its morphology, chromosomes, tumorigenicity and produced a growth curve. CK was detected by immunohistochemistry, the cell cycle determined by flow cytometry and the metastatic potential assessed in 615 and C57BL/c mice. We also transfected the cells with the pEGFP-N1 plasmid. RESULTS A newly established murine cell line was passaged 50 times over a period of 10 months. The cells grow as a partially suspended culture, and are immunohistochemically CK（＋）. The cell line is characterized by a hypotetraploid karyotype, a chromosomal number of 64-68 and a doubling time of 21.8 h. Exponential growth occurs by the third and forth day of culture. Cell cycle analysis showed G1 34%, G2 26%, and 40% in the S phase. The tumorigenicity was 100% upon implantation. No mycoplasma contamination was detected. A monoclonal continuous U14-GFP cell strain which was 100% GFP （＋） was also produced. CONCLUSION We successfully established a new murine cervical U14 carcinoma cell line and an U14-GFP monoclonal strain. These cell lines are ideal for combined in vivo and in vitro tumor research.展开更多
Applephenon®, a purified extract prepared from green apples, was examined for its cytotoxicity and inhibitory effects on the proliferation of cultures of normal human keratinocytes and several epidermoid cancer...Applephenon®, a purified extract prepared from green apples, was examined for its cytotoxicity and inhibitory effects on the proliferation of cultures of normal human keratinocytes and several epidermoid cancer cell lines. Our HPLC studies demonstrated a high content of phenolic compounds (>65%), including catechin, epicatechin, caffeic acid and phloretin as well as polyphenols such as proanthocyanidins. Applephenon® demonstrated a greater cytotoxic effect against HeLa, A431 cancer cell lines and HaCaT, an immortalized keratinocyte cell line than serum-free cultures of proliferating normal human keratinocytes (NHK). Proliferation of NHK was inhibited at concentrations above 0.0013% while concentrations above 0.005% were cytotoxic. By contrast, Applephenon® solutions above 0.00025% killed each of the cancer cell lines. Treated cells displayed increased intercellular separation and evidence of keratinizing stratification. We also tested the effect of epicatechin, and two isoflavonoids, genistein and daidzein, on cancer cell lines. Hela cells were more sensitive to epicatechin and genistein inhibition of cell growth and cytotoxicity than were NHK. Daidzein at these concentrations had little effect on cancer cells. These results indicate that Applephenon® and some of its phenolic components have selective anticancer activity.展开更多
目的:探讨叉头框转录因子M1(FoxM1)对人宫颈癌顺铂耐药株Hela/DDP增殖、侵袭、凋亡的影响及其机制。方法:将Hela/DDP细胞分为空白组(常规培养,不予任何处理)、正常对照(NC)组(转染空白干扰质粒)、低表达组和过表达组。低表达组转染siRNA...目的:探讨叉头框转录因子M1(FoxM1)对人宫颈癌顺铂耐药株Hela/DDP增殖、侵袭、凋亡的影响及其机制。方法:将Hela/DDP细胞分为空白组(常规培养,不予任何处理)、正常对照(NC)组(转染空白干扰质粒)、低表达组和过表达组。低表达组转染siRNA-FoxM1下调Hela/DDP细胞中FoxM1表达,过表达组转染pcDNA3.1-FoxM1上调Hela/DDP细胞中FoxM1表达。采用RT-qPCR法检测FoxM1 m RNA表达水平,CCK-8检测细胞活力,流式细胞仪检测细胞凋亡,Transwell小室检测细胞侵袭能力。结果:FoxM1 mRNA在Hela细胞中的表达水平低于Hela/DDP细胞(P<0.05)。空白组与NC组中FoxM1 mRNA、增殖率、凋亡率、侵袭细胞数及HSP70、S100A9蛋白表达水平比较,差异均无统计学意义(P>0.05)。与空白组比较,低表达组细胞活力和侵袭能力及FoxM1 mRNA、HSP70蛋白表达水平降低,凋亡率和S100A9蛋白表达水平升高(P<0.05),与空白组比较,过表达组细胞活力和侵袭能力及FoxM1 m RNA、HSP70蛋白表达水平升高,凋亡率和S100A9蛋白表达水平降低(P<0.05)。与低表达组比较,过表达组细胞活力和侵袭能力及FoxM1 mRNA、HSP70蛋白表达水平升高,凋亡率和S100A9蛋白表达水平降低(P<0.05)。结论:下调FoxM1表达可抑制HSP70表达,促进S100A9表达,调控宫颈癌细胞恶性行为,从而提高宫颈癌顺铂耐药细胞株对顺铂化疗的敏感性。展开更多
文摘Objective: To discuss the the effect of FGFR4 gene silencing on growth, metastasis and the cis-platinum (DDP) chemotherapy sensitivity of Hela cervical cancer cells. Methods: The Hela cervical cancer cells were cultured and divided into 5 groups: blank control group, negative control group, FGFR4-siRNA group, DDP group and FGFR4-siRNA+DDP group. The siRNA targeted to FGFR4 was constructed and transfected into the cells in FGFR4-siRNA group and FGFR4-siRNA+DDP group, and DDP (10 μg/mL) was added into the DDP group and FGFR4-siRNA+DDP group. The growth inhibition ratio was detected by MTT and the apoptosis rate was detected by FCM after 24 h, 48 h and 72 h culturing respectively. The mRNA and protein expression level of FGFR4, VEGF and MMP2 were detected by RT-PCR and western blot respectively. Results: Compared with the blank control, the proliferative activity of FGFR4-siRNA group decreased after the FGFR4 being silenced, and the growth inhibition ratio and apoptosis rate increased, the FGFR4 did almost not express and the expression level of VEGF and MMP2 decreased.The growth inhibition ratio and apoptosis rate of FGFR4-siRNA+DDP group were higher than that of FGFR4-siRNA group and DDP group, the FGFR4 did almost not express either and the the expression level of VEGF and MMP2 were lower than the FGFR4-siRNA group and DDP group. Conclusion: It can be obtained that silencing the FGFR4 of cervical cancer could inhibit cell growth, promote cell apoptosis and increase the chemosensitivity, down-regulate the expression of VEGF and MMP2 to inhibit the ability of invasion and metastasis, which could be one of the targeted therapeutic targets of cervical cancer.
文摘Objective:To study the inhibitory effect of docetaxel(DOC),oxaliplatin(OXA) and their combination on proliferation of cervical cancer line HeLa.Methods:Cell morphological changes were observed by inverted phase contrast microscope,cell inhibition rates in different groups were examined by MTT,and cell cycle and apoptosis rates were determined by flow cytometry(FCM).Results:DOC and OXA inhibited the proliferation of cervical cancer cell line HeLa in a dose-dependent,and combination of the two drugs had an enhanced inhibitory effect;the apoptosis rate was also significantly increased when the two drugs were used in combination.Conclusion:DOC and OXA can synergistically inhibit the proliferation of cervical cancer cell line HeLa,which indicates that combination of the two drugs might has a promising future for clinical treatment of cervical cancer.
文摘OBJECTIVE To establish a murine uterine cervical cancer cell line and to define its biological characters. METHODS Transplanted tumor tissue was used for in vitro primary culture of U14 cervical carcinoma cells. After 20 passages, we examined its morphology, chromosomes, tumorigenicity and produced a growth curve. CK was detected by immunohistochemistry, the cell cycle determined by flow cytometry and the metastatic potential assessed in 615 and C57BL/c mice. We also transfected the cells with the pEGFP-N1 plasmid. RESULTS A newly established murine cell line was passaged 50 times over a period of 10 months. The cells grow as a partially suspended culture, and are immunohistochemically CK（＋）. The cell line is characterized by a hypotetraploid karyotype, a chromosomal number of 64-68 and a doubling time of 21.8 h. Exponential growth occurs by the third and forth day of culture. Cell cycle analysis showed G1 34%, G2 26%, and 40% in the S phase. The tumorigenicity was 100% upon implantation. No mycoplasma contamination was detected. A monoclonal continuous U14-GFP cell strain which was 100% GFP （＋） was also produced. CONCLUSION We successfully established a new murine cervical U14 carcinoma cell line and an U14-GFP monoclonal strain. These cell lines are ideal for combined in vivo and in vitro tumor research.
文摘Applephenon®, a purified extract prepared from green apples, was examined for its cytotoxicity and inhibitory effects on the proliferation of cultures of normal human keratinocytes and several epidermoid cancer cell lines. Our HPLC studies demonstrated a high content of phenolic compounds (>65%), including catechin, epicatechin, caffeic acid and phloretin as well as polyphenols such as proanthocyanidins. Applephenon® demonstrated a greater cytotoxic effect against HeLa, A431 cancer cell lines and HaCaT, an immortalized keratinocyte cell line than serum-free cultures of proliferating normal human keratinocytes (NHK). Proliferation of NHK was inhibited at concentrations above 0.0013% while concentrations above 0.005% were cytotoxic. By contrast, Applephenon® solutions above 0.00025% killed each of the cancer cell lines. Treated cells displayed increased intercellular separation and evidence of keratinizing stratification. We also tested the effect of epicatechin, and two isoflavonoids, genistein and daidzein, on cancer cell lines. Hela cells were more sensitive to epicatechin and genistein inhibition of cell growth and cytotoxicity than were NHK. Daidzein at these concentrations had little effect on cancer cells. These results indicate that Applephenon® and some of its phenolic components have selective anticancer activity.
文摘目的:探讨叉头框转录因子M1(FoxM1)对人宫颈癌顺铂耐药株Hela/DDP增殖、侵袭、凋亡的影响及其机制。方法:将Hela/DDP细胞分为空白组(常规培养,不予任何处理)、正常对照(NC)组(转染空白干扰质粒)、低表达组和过表达组。低表达组转染siRNA-FoxM1下调Hela/DDP细胞中FoxM1表达,过表达组转染pcDNA3.1-FoxM1上调Hela/DDP细胞中FoxM1表达。采用RT-qPCR法检测FoxM1 m RNA表达水平,CCK-8检测细胞活力,流式细胞仪检测细胞凋亡,Transwell小室检测细胞侵袭能力。结果:FoxM1 mRNA在Hela细胞中的表达水平低于Hela/DDP细胞(P<0.05)。空白组与NC组中FoxM1 mRNA、增殖率、凋亡率、侵袭细胞数及HSP70、S100A9蛋白表达水平比较,差异均无统计学意义(P>0.05)。与空白组比较,低表达组细胞活力和侵袭能力及FoxM1 mRNA、HSP70蛋白表达水平降低,凋亡率和S100A9蛋白表达水平升高(P<0.05),与空白组比较,过表达组细胞活力和侵袭能力及FoxM1 m RNA、HSP70蛋白表达水平升高,凋亡率和S100A9蛋白表达水平降低(P<0.05)。与低表达组比较,过表达组细胞活力和侵袭能力及FoxM1 mRNA、HSP70蛋白表达水平升高,凋亡率和S100A9蛋白表达水平降低(P<0.05)。结论:下调FoxM1表达可抑制HSP70表达,促进S100A9表达,调控宫颈癌细胞恶性行为,从而提高宫颈癌顺铂耐药细胞株对顺铂化疗的敏感性。
