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 研究前列腺癌细胞中miR-221的表达情况及其对癌细胞增殖的影响。方法 运用实时荧光定量PCR(qRT-PCR)检测miR-221在前列腺正常细胞株与前列腺癌细胞株中表达的差异情况,利用细胞转染构建miR-221过表达LNCaP和DU145细胞株,再通过CCK8细胞增殖实验检测细胞增殖情况的变化。结果 qRT-PCR检测细胞株发现miR-221在PC3、LNCaP和DU145三种前列腺癌细胞株中表达量均比前列腺正常细胞株PrEC低 (F=254.197,P<0.001),其中两两比较差异也均有统计学意义。细胞转染技术构建的miR-221过表达LNCaP和DU145细胞株,经qRT-PCR结果显示,miR-221在LNCaP和DU145细胞株中的表达水平明显升高(LNCaP,倍数变化=2.24,t=3.46,P<0.01;Du145,倍数变化=2.24,t=4.29,P<0.01)。细胞增殖实验结果显示,过表达了miR-221的LNCaP(P<0.001)和DU145(P<0.001)细胞生长速度慢于对照组。结论 实验证明miR-221表达过度能减慢前列腺癌细胞的增殖,miR-221有可能成为前列腺肿瘤治疗的生物学标志物。

Objective To investigate miR-221 expression in prostate cancer cells and its influence on prostate cancer cell proliferation. Methods miR-221 expressions in prostate normal cell lines and cancer cell lines were measured by qRT-PCR. Overexpression of the miR-221 in LNCaP and DU145 cell lines used by cell transfection. Effects of the depletion on cell proliferation were assessed in vitro with CCK8. Results qRT-PCR showed miR-221 was lower expressed in PC3, LNCaP and DU145 than in PrEC(F=254.197, P<0.001), in which pairwise comparison also had significant differences. qRT-PCR showed miR-221 expression rose significantly in LNCaP and DU145 cell lines whose miR-221 was overexpression with cell transfection(LNCaP, Fold Change=2.24,t=3.46,P<0.001;Du145, Fold Change=2.24,t=4.29,P<0.001). Cell proliferation assay showed that growth of LNCaP(P<0.001) and DU145(P<0.001) cells whose miR-221 was overexpression was slower than the control group. Conclusion This study demonstrates miR-221 overexpression can inhibited the proliferation of prostate cancer cells for the first time, it also suggests that miR-221 has the potential to serve as a biomarker for PCa therapy.

目的 探讨抗增殖蛋白2（PHB2）脓毒症心肌损伤线粒体功能的调控机制。方法 体外培养大鼠心肌细胞株（H9C2）,分为对照组、脂多糖（LPS）组、LPS+PHB2 siRNA（si-PHB2）组。检测氧化应激指标细胞内丙二醛（MDA）水平、荧光探针检测细胞内活性氧（ROS）水平;线粒体指标：三磷酸腺苷（ATP）水平、线粒体膜电位、线粒体电镜、线粒体半定量评分;免疫印迹法检测PHB2、PTEN诱导激酶1（PTNKI）、帕金蛋白（Parkin）、线粒体转录因子（TFAM）的表达。结果 LPS刺激后MDA水平和ROS水平升高、ATP水平低,LPS+si-PHB2组MDA（6.21±0.39 vs 3.59±0.33, P<0.05）、细胞内的ROS（15 131.37±88.72 vs 8 628.67±71.95, P<0.05）的水平较LPS组升高,ATP（3.46±0.34 vs 4.52±0.25, P<0.05）和线粒体膜电位水平（0.33±0.04 vs 0.55±0.09, P<0.05）进一步降低;电镜观察显示与正常组相比,LPS组、LPS+si-PHB2组出现不同程度线粒体损伤,线粒体损伤半定量评分显示LPS+si-PHB2组的损伤较LPS组更为明显（1.42±0.10 vs 0.81±0.04, P<0.05）; 免疫印迹法结果显示LPS处理后PHB2、PINK1、Parkin 表达上调,TFAM表达下调,LPS+si-PHB2组的线粒体自噬相关蛋白PINK1（1.33±0.06 vs 1.79±0.21, P<0.05）、Parkin（1.43±0.08 vs 1.86±0.09, P<0.05）和线粒体生物发生关键蛋白TFAM（0.29±0.01 vs 0.74±0.06, P<0.05）表达均较LPS组降低。结论 LPS可促进大鼠心肌细胞PHB2表达,si-PHB2干扰后线粒体自噬蛋白和生物发生蛋白表达抑制,心肌细胞氧化应激损害和线粒体功能障碍加重,提示PHB2表达上调可能恢复线粒体稳态改善脓毒症心肌损伤的线粒体功能。

Objective To explore the regulatory mechanism of septic myocardial injury by prohibitin 2（PHB2）. Methods Rat myocardial cell lines（H9C2）were cultured in vitro and divided into control group,LPS group,LPS + PHB2 siRNA（si-PHB2） group. The indicators for detecting oxidative stress include the levels of intracellular malondialdehyde（MDA）and reactive oxygen species（ROS）. The indicators for mitochondrial detection include adenosine triphosphate（ATP）levels,mitochondrial membrane potential,mitochondrial electron microscopy,and semi-quantitative mitochondrial scoring. Western blotting was used to detect the expression of PHB2,PTEN induced putative kinase（PINK1）,Parkin,mitochondrial transcription factor A（TFAM）. Results After LPS stimulation,MDA level and intracellular ROS level increased,ATP level decreased. Compared with LPS group,MDA（6. 21±0. 39 vs 3. 59±0. 33, P<0. 05）level and intracellular ROS level（15 131. 37±88. 72 vs 8 628. 67±71. 95, P<0. 05）in LPS + si-PHB2 group increased significantly,while ATP（3. 46±0. 34 vs 4. 52±0. 25, P<0. 05）and MMP（0. 33±0. 04 vs 0. 55±0. 09, P<0. 05）level further decreased. Compared with the normal group,the structure of mitochondria in LPS group and LPS + si-PHB2 group was damaged in different degree. The semi-quantitative score of mitochondrial damage showed that the damage in LPS + si-PHB2 group was more obvious than that in LPS group（1. 42±0. 10 vs 0. 81±0. 04, P<0. 05）. Western blotting showed that the expression of PHB2,PINK1 and Parkin were up-regulated and the expression of TFAM was down-regulated after LPS treatment,mitohagy-related proteins PINK1（1. 33±0. 06 vs 1. 79±0. 21, P<0. 05）,Parkin（1. 43±0. 08 vs 1. 86±0. 09, P<0. 05）and mitochondrial biogenetic protein TFAM（0. 29±0. 01 vs 0. 74±0. 06, P<0. 05）in LPS+si-PHB2 group were lower than those in LPS group. Conclusions LPS can promote the expression of PHB2 in rat cardiomyocytes. After interfering with PHB2 expression,we found that mitochondrial autophagy and biogenesis are inhibited,and mitochondrial dysfunction,oxidative stress exacerbated,suggesting that the up-regulation of PHB2 expression may restore mitochondrial homeostasis and improve mitochondrial function in septic myocardial injury.

       目的   研究SOCS6/STAT6通路在前列腺细胞炎性增殖作用中的调控作用。方法  使用人前列腺细胞株RWPE-1建立炎症模型，将细胞分为对照（CON）组和炎症刺激（INF）组，后者通过添加脂多糖（LPS）模拟炎症环境。采用ELISA检测白细胞介素-1β（IL-1β）-1β、白细胞介素-6（IL-6）和白细胞介素-8（IL-8）表达水平，蛋白免疫印迹法检测细胞因子信号抑制物-6（SOCS6）、信号转导和转录激活因子-6（STAT6）及磷酸化STAT6蛋白的表达水平。结果  经过LPS处理后，RWPE-1细胞中的SOCS6蛋白表达水平显著下降（P＜0.01），而磷酸化STAT6表达水平上升（P＜0.01）。结论  SOCS6/STAT6通路可能通过调节炎症环境下STAT6的磷酸化水平，参与调节前列腺细胞的炎性增殖作用。

       Objective   To explore the  regulatory  role of  SOCS6/STAT6  pathway in the inflammatory  proliferation of 

prostate cells．Methods  The human prostate cell line RWPE-1 was used to establish an inflammation model．Cells were divided into a control（CON）group and an inflammation-stimulated（INF）group，with the latter subjected to lipopolysaccharide（LPS）treatment to simulate an inflammatory environment．The expression levels of interleukin（IL）-1β、IL-6 and  IL-8 were detected by ELISA，and the expression levels of suppressor of cytokine signaling 6（SOCS6），signal transducer and activator oftranscription-6，（STAT6），and phosphorylated STAT6 proteins were detected by Western blot．Results  The  results showed that after LPS treatment，the expression of SOCS6 protein in RWPE-1 cells significantly decreased，while the expression of phosphorylated STAT6 increased．Conclusions  The SOCS6/STAT6 pathway may be involved in  regulating the inflammatory proliferation of prostate cells by modulating the phosphorylation level of STAT6 under inflammatory conditions．