目的 探讨不同浓度的青藤碱对肾癌细胞增殖、细胞周期及凋亡的影响。方法 以不同浓度的青藤碱处理肾癌细胞786-O,采用四氮唑蓝盐法检测细胞增殖能力,流式细胞术检测细胞周期分布,Annexin-v FITC/PI双染流式细胞分析仪检测细胞凋亡率;采用实时荧光定量PCR及蛋白免疫印迹(WB)检测c-myc、Bax、Caspase-3等细胞周期、凋亡相关基因表达情况。结果 青藤碱显著抑制786-O细胞的增殖能力,诱导细胞周期G1/S期阻滞及细胞凋亡;且随着青藤碱浓度的增加,其抑制率也逐渐增加;青藤碱显著下调c-myc蛋白表达,而诱导凋亡蛋白Bax、Caspase-3表达上调。结论 青藤碱可以显著抑制786-O细胞中c-myc表达,使其增殖能力减弱,诱导细胞周期阻滞及凋亡。青藤碱可能具有潜在的抑制肾癌生长作用。

Objective To investigate the effects of different concentrations of sinomenine on proliferation, cell cycle and apoptosis of renal carcinoma cells. Methods The renal carcinoma cells were treated with different concentrations of sinomenine. MTS was used to analyze the effects of sinomenine on proliferation in 786-O renal carcinoma cells, the cell cycle changes were determined using flow cytometry, while the changes of apoptosis were detected by Annexin V-FITC / PI double staining. The expression of apoptosis-related proteins such as c-myc, Bax and Caspase-3 were detected by Western blot. Results Sinomenine significantly inhibited the proliferation of 786-O cells and induced cell cycle arrest and apoptosis in G1/S phase. With the increase of sinomenine concentration, the inhibition rate increased gradually. Sinomenine significantly down-regulated the expression of c-myc protein, while the expressions of the apoptotic protein Bax, Caspase-3 were up-regulated. Conclusions Sinomenine can significantly inhibit the expression of c-myc in 786-O cells, reduce proliferation ability, and induce cell cycle arrest and apoptosis. Sinomenine may have a potential therapeutic effect on renal cancer.

目的 利用分析各种浓度环氧化酶-2(COX-2)特异度抑制剂塞来昔布对食管癌EC109细胞系的作用,进而对COX-2蛋白表达的影响及对细胞凋亡能力的作用,进一步探讨塞来昔布对食管癌细胞凋亡的作用及机制。方法 使用0 μmol/L、20 μmol/L、60 μmol/L、100 μmol/L四个浓度的塞来昔布处理EC109细胞24 h,酶联免疫吸附剂测定(ELISA)法测定COX-2蛋白表达;流式细胞仪测定EC109细胞凋亡情况。结果 与0 μmol/L塞来昔布组比较,20 μmol/L、60 μmol/L、100 μmol/L塞来昔布组EC109细胞内COX-2蛋白表达不断降低(1.581±0.116;1.226±0.089,0.846±0.076,0.521±0.082)(P<0.05);而细胞凋亡率逐步上升(1.700±0.557,13.400±1.735,18.766±1.301,28.100±1.997)(P<0.05)药物浓度依赖于梯度。结论 塞来昔布是一种COX-2抑制剂,可能以浓度梯度的形式抑制COX-2蛋白的表达,从而促进EC109细胞的凋亡。

Objective The effects of celecoxib, a specific COX-2 inhibitor at various concentrations, on EC109 cell line of esophageal cancer were analyzed, and the effect and mechanism of celecoxib on apoptosis of esophageal carcinoma cells were further studied. Methods EC109 cells were treated with celecoxib at concentrations of 0 μmol/L, 20 μmol/L, 60 μmol/L and 100 μmol/L for 24 h. The protein of COX-2 in EC109 cells was determined by enzyme-linked immunosorbent assay (ELISA). Assay of EC109 cell apoptosis were determined by flow cytometry. Results Compared with the 0μmol/L celecoxib group, the expression of COX-2 protein in EC109 cells of 20μmol/L, 60μmol/L, 100μmol/L celecoxib group gradually decreased(1.581±0.116; 1.226±0.089, 0.846± 0.076, 0.521±0.082) (P<0.05); and the apoptotic rate gradually increased (1.700±0.557; 13.400±1.735, 18.766±1.301, 28.100±1.997) (P<0.05) in a drug concentration gradient-dependent manner. Conclusion The COX-2 inhibitor celecoxib may inhibit the expression of COX-2 protein in a concentration gradient and promote the apoptosis of esophageal cancer EC109 cells.

目的 探讨细胞毒素-1(Cytotoxin-1,CTX-1)对人卵巢癌SKOV-3细胞增殖凋亡的影响。方法 利用0、4、8、12 μg/mL浓度 CTX-1处理SKOV-3细胞6、12、24 h,MTS法检测细胞活性,8 μg/mL CTX-1处理SKOV-3细胞24、48 h,Hoechst-33258荧光染色观察细胞核染色质形态。取处理 6、12 h 后细胞,利用流式细胞仪检测SKOV-3细胞的凋亡率。结果 4、8、12 μg/mL的CTX-1可抑制SKOV-3细胞活性及增殖,呈时间-剂量依赖。Hoechst-33258染色观察可见细胞染色质呈固缩或碎裂状、染色质着色不均、核形态各异,随时间增加而更趋明显。8 μg/mL CTX-1处理细胞,6 h细胞坏死率为(1.90±0. 27)%,晚期凋亡率为(10.96±1. 56)%,而早期凋亡率为(1.52±0.39)%;12 h细胞坏死率为(10.62±0.96)%,晚期凋亡率(15.07±1.23)%,而早期凋亡率为(1.88±0.17)%,与对照组比较,差异有统计学意义 (P<0.0 1)。结论 CTX-1可以抑制人卵巢癌细胞活性、抑制其体外增殖、诱导其发生凋亡,该作用呈剂量依赖和时间依赖,主要引起细胞晚期凋亡和坏死。

Objective To investigate the effect of cytotoxin-1 (CTX-1)on the proliferation and apoptosis of ovarian cancer SKOV-3 cells. Methods SKOV-3 cells were treated with CTX-1 at concentrations of 0, 4, 8, 12 μg/mL for 6, 12, and 24 hours respectively. Cell viability was measured by MTS method. SKOV-3 cells were treated with 8 μg/mL CTX-1 for 24 and 48 hours, by Hoechst-33258 fluorescence staining to observe the morphology of nuclear chromatin. The apoptotic rate of SKOV-3 cells was detected by flow cytometry after 6 and 12 hours of treatment. Results CTX-1 at 4, 8, and 12 μg/mL inhibited the activity and proliferation of SKOV-3 cells in a time-dose-dependent manner. Hoechst-33258 staining observation showed that the apoptotic cell chromatin was condensed or fragmented chromatin, the chromatin was unevenly colored, and the nuclear morphology was different. It became more obvious with time. 8 μg/mL CTX-1 treated cells, the 6 h cell necrosis rate was (1.90±0.27)%, the late apoptosis rate was (10.96±1.56)%, and the early apoptosis rate was (1.52±0.39)%; 12 hours cell necrosis rate was (10.62±0.96)%, late apoptosis rate was (15.07±1.23)%, and early apoptosis rate was (1.88±0.17)%, compared with the control group, the difference was statistically significant (P<0.01). Conclusion CTX-1 may inhibit the activity of human ovarian cancer cells, inhibit its proliferation in vitro, and induce its apoptosis. The effect is dose-dependent and time-dependent. Mainly it causes late apoptosis and necrosis of cells.

目的 在原来研究的基础上进一步研究Wnt-1信号通路蛋白-3（WISP-3）在高氧诱导肺上皮细胞凋亡中的作用。方法 通过Western blot检测和免疫组化检测不同肺上皮细胞中WISP-3的蛋白表达量。利用质粒转染和siRNA的方法在Beas-2B细胞中高表达和基因沉默WISP-3,通过细胞活性检测和流式细胞学技术检测高氧刺激后细胞的凋亡情况。结果 与空气对照相比,高氧刺激使肺上皮细胞的WISP-3蛋白表达量下降;WISP-3基因沉默或高表达使高氧诱导的肺上皮细胞凋亡增加或减少。结论 高氧刺激下,肺上皮细胞中WISP-3表达下降,WISP-3对高氧诱导的肺上皮细胞凋亡具有保护作用。

Objective To explore how Wnt-1 inducible signaling pathway protein-3 （WISP-3） participate in and play a regulatory role in the process of hyperoxia induced apoptosis in lung epithelial cells. Methods The expression of WISP-3 was detected via Western blot and immunohistochemistry. High expression and low expression of WISP-3 were performed by plasmid transfection and siRNA. Cell viability and flow cytometry were executed to detect the hyperoxia-induced apoptosis in Beas-2B. Results Compared to the group of air control,the expression of WISP-3 protein in lung epithelial cells decreased obviously after hyperoxia. Cell survival decrease and apoptosis increased after hyperoxia in Beas-2B cells with low expression of WISP-3. Vice versa. Conclusion The expression of WISP-3 decreased after hyperoxia in lung epithelial cells. The role of WISP-3 in this process may be protective.

目的 探讨Reversine对人肝星状细胞系LX-2凋亡的影响。方法 设对照组和Reversine干预组,其中Reversine干预组分为7个浓度,分别为1,5,10,20,40,80,120 μg/mL,CCK-8法检测Reversine对LX-2增殖的影响,选取最佳浓度。将细胞重悬在加入5 μL FITC-Annexin V和5 μL PI,用流式细胞仪进行了凋亡率分析,免疫荧光检测凋亡蛋白bcl-2及caspase 3。结果 Reversine可促进LX-2细胞凋亡,随着Reversine浓度增加,LX-2的凋亡可呈剂量依赖关系,其中10 μg/mL为最佳浓度,LX-2细胞的bcl-2蛋白的表达显著下降而cleaved-caspase 3的表达显著上升。结论 Reversine可通过促进caspase-3蛋白活化、抑制bcl-2蛋白表达的方式诱导LX-2凋亡。

目的 探索蟾毒灵对舌鳞状细胞癌Tca8113细胞增殖、凋亡的影响及可能作用机制。方法 以人舌鳞癌Tca8113细胞为研究对象,MTT法检测10、20、40、80、160 nmol/L浓度蟾毒灵体外抑制Tca8113细胞增殖的活性;检测蟾毒灵干预下肿瘤细胞Na⁺-K⁺-ATP酶活性的变化;Western blot发检测Bcl-2、Bax、caspase-3蛋白表达。结果 蟾毒灵有抑制Tca8113细胞的活性,且呈剂量-时间依赖性;在蟾毒灵干预下Tca8113细胞Na⁺-K⁺-ATP酶收到抑制;Western blot结果显示凋亡相关Bax、caspase-3蛋白表达上调,Bcl-2蛋白表达下调。结论 蟾毒灵通过抑制细胞膜Na⁺-K⁺-ATP酶活性,通过调节Bcl-2凋亡通路的相关蛋白,最终激活caspase-3,诱导人舌鳞癌Tca8113细胞凋亡。

Objective To explore the effects of bufalin on proliferation and apoptosis of tongue squamous cell carcinoma Tca8113 cells and its possible mechanism. Methods Tca8113 cells were treated with 10, 20, 40, 80 and 160 nmol/L Tca8113 cells in vitro. MTT assay was used to detect the inhibitory effect of bufalin on the proliferation of Tca8113 cells; And the activity of Na⁺-K⁺-ATPase in tumor cells was detected by the interference of bufalin; The expression of Bcl-2, Bax and caspase-3 protein was detected by Western blot. Results Bufalin inhibited the activity of Tca8113 cells in a dose-and time-dependent manner; Na⁺-K⁺-ATPase in Tca8113 cells was inhibited by bufalin; The results of Western blot showed that the expression of Bax and caspase-3 protein was up-regulated and the expression of Bcl-2 protein was down-regulated. Conclusion Bufalin induced the apoptosis of human tongue squamous cell carcinoma Tca8113 cells by inhibiting the activity of Na⁺-K⁺-ATPase and regulating the related proteins of Bcl-2 apoptosis pathway, finally activating caspase-3.

目的 观察紫河车提取物联合顺铂对人脑胶质瘤细胞增殖与凋亡的影响。方法 把正常培养传代后的U251胶质瘤细胞按随机分配的方法分为四组,A组仅加普通培养液,B、C、D组各加紫河车提取物(400 mg/mL)2 mL、顺铂(1 mg/mL)0.01 mL、紫河车提取物(400 mg/mL)2 mL＋顺铂(1 mg/mL)0.01 mL;MTT法观察U251细胞增殖情况,流式细胞仪检测U251细胞凋亡率。结果 培养12、24、36、48、60 h,B、C、D组细胞增殖指数逐渐下降,与A组进行比较,各组P值均小于0.05;其中,将D组与B、C组进行比较,P值小于0.05。将各组培养24 h后上机,测得A、B、C、D各组细胞的凋亡率分别为(0.3±0.2)%,(10.6±1.5)%,(35.9±2.8)%,(52.1±4.1)%。其中,B、C、D各组和A组进行比较,P值均小于0.05;将D组与B、C组两组进行比较,P值也均小于0.05。结论 紫河车提取物联合顺铂可抑制人脑胶质瘤U251细胞增殖,并诱导其凋亡。

Objective To observe the effect of cisplatin combinated with the placental immunoregulating polypeptide (PIP) on proliferation and apoptosis of glioma cells. Methods Randomly we divide the normal handed U251 glioma cells into four groups. We added ordinary nutrient solution to group A, while added activated PIP(400 mg/mL)2 mL to group B, cisplatin (1 mg/ml) 0.01 ml to group C, PIP 400 mg/mL)2 mL and cisplatin (1 mg/mL) 0.01 mL to group D. We surveyed the proliferation rate of gliobma cells by MTT experimental method and analyzed the apoptosis of U251 glioma cells by flow cytometry. Results The index of cell proliferation of group B,C,D declined gradually with the training of 12 h,24 h,36 h,48 h,60 h. Compared B, C,D group with A group, P<0.05,and compared group D with group B and group C, P< 0.05. Put groups culturing of 24 hour on flow cytometer, the glioma cells apoptosis rate of each group was 0.3%±0.2%、10.6%±1.5%、35.9%±2.8%、52.1%±4.1% respectively. Compared group B,C,D with group A, P<0.05,and compared group D with group B and group C, P<0.05. Conclusion Placental immunoregulatingpPolypeptide combined with cisplatin may restrain the proliferation of human glioma cells, meanwhile increase the apoptosis of glioma cells.

目的 氧化苦参碱对视网膜母细胞瘤细胞SM-106凋亡的诱导作用及机制。方法 以不同作用时间(24 h、48 h、72 h)和不同作用浓度(12.5 μl/mL、25 μl/mL、50 μl/mL、100 μl/mL)氧化苦参碱处理视网膜母细胞瘤细胞SM-106,分别采用流式细胞仪及western blot检测视网膜母细胞瘤细胞SM-106细胞凋亡及其凋亡因子(Bax、Bcl-2)蛋白表达。结果 氧化苦参碱可促进SM-106细胞体外凋亡,上调Bax蛋白表达及Bax/Bcl-2蛋白表达比值,下调Bcl-2蛋白表达,并呈现剂量及时间依赖性。结论 氧化苦参碱可诱导视网膜母细胞瘤细胞SM-106凋亡,调控凋亡因子Bax、Bcl-2的表达是其可能作用机制。

Objective To evaluate the apoptosis and its mechanism of retinoblastoma cells SM-106 induced by oxymatrine. Methods Retinoblastoma cells SM-106 were treated with different time(24 h、48 h、72 h)and different concentrations(12.5 μl/mL, 25 μl/mL, 50 μl/mL or 100 μl/mL) of oxymatrine. The apoptosis and protein expression of apoptosis factors (Bax and Bcl-2) were respectively determined by flow cytometry and western blot. Results Oxymatrine significantly promoted the SM-106 cells apoptosis in vitro, raised Bax protein expression and Bax/Bcl-2 protein expression ratio, reduced the Bcl-2 protein expression, and showed the dose and time dependent. Conclusion Oxymatrine is able to induce the apoptosis in retinoblastoma cells SM-106. Regulating apoptosis related gene Bax and Bcl-2 expression may be the mechanism of apoptosis.

目的 观察葛根素对新西兰白兔视网膜缺血/再灌注损伤组织中细胞凋亡的保护作用。方法 新西兰白兔30只随机分为缺血再灌注对照组和葛根素治疗实验组,各组右眼应用前房灌注加压法建立视网膜缺血再灌注模型,分别于再灌注后第12、24、72 h处死动物,摘除眼球,制作石蜡切片,用TUNEL法检测细胞凋亡,计算凋亡指数。结果 对照组缺血再灌注12 h在神经节细胞层和内核层可见凋亡细胞;24 h神经节细胞层细胞数有所减少,视网膜神经节细胞层、内核层及外核层均见凋亡细胞明显增多;72 h神经节细胞层细胞数明显减少,神经节细胞层、内核层及外核层仍见凋亡细胞,但较24 h有所减少。葛根素治疗视网膜的凋亡细胞在各个时间段的表达规律与对照组相似,但凋亡细胞计数在12 h,24 h,72 h均较对照组明显减少,两组间差异有统计学意义。结论 葛根素能减轻缺血-再灌注损伤的视网膜细胞凋亡,对视网膜有保护作用。

Objective To observe the protective effects of Puerarin on apoptosis of ischemic injury in rabbit retina. Methods Retinal ischemia was induced in rabbits by increasing intraocular pressure to 120 mmHg for 60 minutes. TdT-mediated biotin-dUTP nick end labelling(TUNEL) staining technique was used to examine the apoptosis of retinal ganglion cells in the control group and the puerarin treatment group. Results The number of apoptotic cells in 12, 24 and 72h after reperfusion in the puerarin treatment group was obvious lower than that in the control group(P<0.05). Conclusion Puerarin has protective effects in protecting against apoptosis in ischemia reperfusion injury of rabbit retina.