野艾蒿挥发油对HeLa癌细胞形态与结构的影响

采用MTT法检测细胞活力,用倒置显微镜、荧光显微镜和扫描电子显微镜观察细胞形态与结构的变化,用激光共聚焦显微镜观察细胞微管的分布,从而研究了野艾蒿挥发油对HeLa人宫颈癌细胞形态与结构的影响。结果表明:(1)野艾蒿挥发油对HeLa癌细胞的增殖有明显的抑制作用,呈剂量和时间依赖性。(2)野艾蒿挥发油处理HeLa癌细胞24h后,100、200μg/mL实验组细胞体积缩小,核染色质凝集、微绒毛消失、细胞表面有泡状突起,微管解聚,呈现典型的凋亡特征;400μg/mL实验组细胞膜破裂、胞浆内含物外泄,呈明显的坏死特征。(3)野艾蒿挥发油具有抑制HeLa癌细胞增殖的作用,低、中浓度的野艾蒿挥发油诱导细胞凋亡,而高浓度的野艾蒿挥发油引起细胞坏死。

野艾蒿挥发油诱导HeLa细胞凋亡与坏死

目的:研究野艾蒿挥发油对HeLa细胞凋亡与坏死的影响。方法:MTT法检测细胞活力,显微镜观察细胞形态与结构的变化,DNA Ladder、流式细胞仪检测细胞凋亡,彗星电泳检测DNA损伤,Western blot检测Caspase-3和多聚ADP核糖聚合酶(PARP)表达。结果:MTT法检测结果显示野艾蒿挥发油对HeLa细胞的增殖有明显的抑制作用,呈剂量依赖性。野艾蒿挥发油处理HeLa细胞24 h后,100、200μg/mL实验组细胞体积缩小,核固缩,染色质凝集、片段化,表现为典型的凋亡特征;400μg/mL实验组细胞膜破裂、内含物外泄,表明细胞已坏死。此外,DNA Ladder结果显示,各实验组均有梯状条带;流式细胞仪检测结果也显示,各实验组均有凋亡峰。野艾蒿挥发油处理HeLa细胞6 h后,100μg/mL实验组彗星电泳中出现彗尾。Western-blotting结果显示,野艾蒿挥发油能够诱导HeLa细胞中Caspase-3蛋白表达量降低,同时PARP发生剪切、失活。结论:野艾蒿挥发油具有抑制HeLa细胞增殖的作用。低浓度野艾蒿挥发油诱导细胞凋亡,高浓度野艾蒿挥发油引起细胞坏死,其凋亡机制是依赖Caspase-3介导的PARP凋亡信号转导途径。

碳源饥饿对癌细胞和正常细胞增殖的影响

研究了共培养条件下碳源饥饿对癌细胞和正常细胞形态、生长和凋亡的影响.采用细胞体外共培养的方法,实验分4组:对照组(25 mmol/L葡萄糖)及5,1和0 mmol/L葡萄糖组,分别处理共培养条件下的HeLa癌细胞和人脐带正常细胞,在倒置显微镜下观察细胞的形态与生长,吖啶橙/溴乙锭(AO/EB)双重荧光染色检测凋亡细胞的形态及数量.结果表明:对照组HeLa癌细胞和人脐带正常细胞都分布均匀,贴壁性好,形态饱满,多呈规则的梭形或多角形,折光性好,AO/EB双染绿色荧光分布均匀;实验组糖浓度越低,细胞存活时间越短,细胞收缩变圆并逐渐凋亡,AO/EB双染细胞由绿色荧光逐渐变为橙黄色荧光;相同糖浓度条件下人脐带正常细胞比HeLa癌细胞存活时间更长.说明碳源饥饿使癌细胞先于正常细胞死亡.

Effects of antisense oligonucleotides targeting VEGF on radio sensitivity of uterine cervix cancer Hela cells

Objective： To determine the impact of antisense oligonucleotides targeting vascular endothelial growth factor （VEGF） on radiosensitivity of uterine cervix cancer Hela cells. Methods： VEGF antisense oligodeoxynucleotides （ASODN） was transfected into Hela cells by liposome-mediated method. Cells transfected with the oligodeoxynuclecotide and saline were used as control groups. Cells were irradiated by 6 MV X ray at the dose of 0 Gy, 2 Gy, 4 Gy and 6 Gy respectively. The expression of VEGF mRNA was determined by RT-PCR. Apoptosis were evaluated using FCM. Cloning efficiency was determined by colony formation assay. Results： The expression of VEGF mRNAwas inhibited by ASODN （P 〈 0.01） in Hela cells. The inhibited activation which was influenced by radiation resulted in increasing apoptosis （P 〈 0.01） and inhibiting plating efficiency （P 〈 0.01）. Conclusion： The expression of VEGF induced by X irradiation in Hela cells can be blocked by VEGF ASODN. Treatment with VEGF might increase apoptosis in HeLa cells and enhance radiosensitivity.

Effects of docetaxel,oxaliplatin and their combination on HeLa strain of cervical cancer

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.

Effect of aspirin alone or combined with cisplatin on human cervical carcinoma HeLa cells

Objective： To study the effect and mechanism of aspirin alone or combined with cisplatin （DDP） on human cervical carcinoma HeLa cells. Methods： HeLa cells were treated by different concentrations of aspirin, DDP alone or both. The inhibitory effect on cell growth was analyzed by MTT and colony-forming assay. Cell apoptosis was measured with flow cytometry. The mRNA levels ofBcl-2, Bax and NF-κB（P65） were studied by RT-PCR. Results： MTT assay showed that aspirin inhibited HeLa cell proliferation in a time-and dose-dependant maoner. Aspirin decreased clone numbers in colony formation assay. Aspirin also induced apoptosis of HeLa cells in a dose- and time-dependent manner as detected by flow cytometry. The inhibition effects on proliferation, colony formation and apoptosis were significantly enhanced when cells were treated with both aspirin and DDP. RT-PCR demonstrated that aspirin decreased the transcription of Bcl-2 and NF-κB, and increased expression of Bax gene. Conclusion： Aspirin can induce apoptosis in HeLa cells. Combination of aspirin and DDP displays a synergistic effect. The possible mechanism might be that aspirin downregulates the mRNA levels of Bcl-2 and NF-κB gene and upregulates the expression of Bax.

大气压冷等离子体射流灭活子宫颈癌Hela细胞

研究了大气压冷等离子体射流对子宫颈癌Hela细胞的灭活机制.在倒置显微镜下观察不同等离子体处理条件下的细胞形态,并通过中性红吸收测试定量测定各个条件下的细胞存活率.将功率维持在18W,在900mL/min氩等离子体中添入氧气的百分含量分别为1%,2%,4%和8%的条件下处理Hela细胞,探讨活性气体氧气在惰性气体氩气中的百分含量对Hela癌细胞灭活效率的影响,发现添加2%氧气时,氩/氧等离子体灭活效果最佳,处理180s后细胞存活率可降至7%.当继续添加氧超过2%时,灭活效果逐渐减弱,直至8%时,其效果反而不如单纯氩等离子体.通过测量等离子体发射光谱,结果表明活性氧自由基在癌细胞灭活过程中可能起关键作用.

藻蓝蛋白的抗癌活性研究

活细胞脱氢酶催化四唑盐 (MTT)还原 ,生成溶于有机溶剂的蓝色甲月赞结晶 ,可定量反映细胞生长 .流式细胞技术可根据细胞中 DNA的质量分数来检测其所处的细胞周期 .采用上述两种方法 ,研究了藻蓝蛋白 (PC)对 He La细胞的抗癌活性 .实验证明 ,PC对 He La细胞生长有明显的抑制作用 ,当 PC浓度为 80 mg/L时对癌细胞的抑制率达 31.0 %.初步认为这种抑制机理为 :PC使 He L a细胞由合成期(S)或分裂期 (M)向 G1期转变和积集 ,细胞 DNA合成衰减 .

Expression of PADI4 in hepatocellular carcinoma

Objective： The aim of the research was to study peptidylarginine deiminase type 4 （PAD4/PADI4） expression and its tumodgenic mechanism in hepatocellular carcinomas. Methods： Expressions of PADI4 and p53 were investigated in tumors and non-tumor tissues by Western blot in patients with hepatocellular carcinomas. We constructed plasmid of PADI4-Flag and transfected it in Hela cells to investigate the mechanism. Results： Western blot analysis showed higher PADI4 expression in hepatocellular carcinomas than in the surrounding healthy tissues. Furthermore, by Western blot, we detected decreased p53 levels in the tumor tissues of patients with hepatocellular carcinomas compared to surrounding healthy tissues. In Hela cells transfected with PcDNA3.0-Flag-PADI4 plasmid, the expression of p53 decreased obviously. Conclusion： Our results suggest that PADI4 elevated in the tissues of hepatocellular carcinomas and induced tumorigenic by down-regulating p53 expression.

Inhibition of Aspergillus parasiticus and Cancer Cells by Marine Actinomycete Strains

Ten actinomycete strains isolated from the Yellow Sea off China＇s coasts were identified as belonging to two genera by 16S rDNA phylogenetic analysis： Streptomyces and Nocardiopsis. Six Streptomyces strains （MA10, 2SHXF01-3, MA35, MA05-2, MA05-2-1 and MA08-1） and one Nocardiopsis strain （MA03） were predicted to have the potential to produce aromatic polyketides based on the analysis of the KSa （ketoacyl-synthase） gene in the type II PKS （polyketides synthase） gene cluster. Four strains （MA03, MA01, MA10 and MA05-2） exhibited significant inhibitory effects on mycelia growth （inhibition rate 〉50%） and subsequent aria- toxin production （inhibition rate 〉75%） of the mutant aflatoxigenic Aspergillus parasiticus NFRI-95. The ethyl acetate extracts of the broth of these four strains displayed significant inhibitory effects on mycelia growth, and the IC50 values were calculated （MA03： 0.275 mg mL-1, MA01：0.106 mg mL-1, MA10：1.345 mg mL-1 and MA05-2：1.362 mg mL-1）. Five strains （2SHXF01-3, MA03, MA05-2, MA01 and MA08-1） were selected based on their high cytotoxic activities. The ethyl acetate extract of the Nocardiopsis strain MA03 was particularly noted for its high antitumor activity against human carcinomas of the cervix （HeLa）, lung （A549）, kidney （Caki-1） and liver （HepG2） （IC50： 2.890, 1.981, 3.032 and 2.603 μgmL-1, respectively）. The extract also remarkably inhibited colony formation of HeLa cells at an extremely low concentration （0.5μgmL 1）. This study highlights that marine-derived actinomycetes are a huge resource of compounds for the biological control of aflatoxin contamination and the development of novel drugs for human carcinomas.

Up-regulation of Raf kinase inhibitor protein enhances chemosensitivity of cervical cancer cell

Objective： The purpose of the study is to investigate the effects of up-regulation of Raf kinase inhibitor protein （RKIP） on the chemosensitivity of cervical cancer Hela cells. Methods： Eukaryotic expression plasmid pcDNA3.1（±）-ssRKIP containing human overall length RKIPcDNA was transfected into cervical cancer Hela cell by lipofectin assay, establishing a stable cell line containing a target gene by G418. Expression of RKIP in Hela cells was measured by Western blot analysis. After treatment with cisplatin of different concentrations and intervals of time, the effect of RKIP on the proliferation of Hela cells was evaluated by MTT method. The flow cytometry was used to investigate whether the RKIP could inhibit apoptosis in Hela cells induced by cisplatin. Results： The expression of RKIP in Hela cells transfected with pcDNA3.1-ssRKIP was increased obviously. After different concentrations of cisplatin treatment cells for 24, 48 and 72 h, the growth inhibition rate in Hela cells transfected with pcDNA3.1-ssRKIP was significantly higher than in control cells （P 〈 0.05）. With 5 pg/mL cisplatin treatment for 24 h, pcDNA3.1-ssRKIP-transfected Hela cells had an obviously higher percentage of apoptosis （23.2 ± 0.24）% than non-transfected cells （12.4 ± 0.31）% and empty vector-transfected cells （13.4 ± 0.47）%. Without treatment of cisplatin, the percentage of apoptosis for Hela cells transfected with pcDNA3.1-ssRKIP was （5.7 ± 0.12）%, which was still higher than those of the non-transfected cells （2.9 ± 0.21）% and empty vector-transfected cells （3 ± 0.08）%. Conclusion： Higher expres- sion of RKIP gene can improve chemosensitivitv of cervical cancer Hela cells to cisplatin.