目的 探讨长链非编码核糖核酸肺腺癌转移相关转录本 1（LncMALAT1）通过竞争性结合微小RNA-506-3p（miR-506-3p）调控Zeste同源物增强子2（EZH2）影响膀胱癌增殖的机制。方法 收集2023年1月—2024年10月的92例外科手术切除的膀胱癌组织及对应的癌旁组织标本, 利用Western blot和定量实时逆转录聚合酶链式反应（qRT-PCR）方法检测LncMALAT1和EZH2的表达情况。根据患者预后分为不良组（n=34）和良好组（n=58）, 收集患者的性别、年龄、肿瘤直径、血管侵袭情况、TNM分期、远处转移情况等临床指标, 结合临床病理指标分析LncMALAT1和EZH2与膀胱癌患者预后的关系。通过体外实验,包括qRT-PCR、Western blot、平板克隆和EdU实验,验证LncMALAT1对EZH2表达和膀胱癌细胞增殖的影响。利用生物信息学技术预测LncMALAT1与miR-506-3p的相互作用,并通过qRT-PCR验证在膀胱癌细胞中上调LncMALAT1表达后miR-506-3p的表达变化。结果 单因素结果显示, 血管侵袭情况、TNM分期、远处转移情况、LncMALAT1及EZH2表达水平均与膀胱癌患者预后不良有关, 差异有统计学意义（均P<0.05）。分析结果发现LncMALAT1与EZH2在膀胱癌组织中的表达呈正相关。体外实验结果显示, 上调LncMALAT1表达后, EZH2的表达显著上调, 且膀胱癌细胞的增殖能力显著提高（均P<0.05）。qRT-PCR验证表明,上调LncMALAT1表达后,miR-506-3p的表达显著下调（P<0.05）, 提示LncMALAT1通过竞争性结合miR-506-3p调控EZH2,进而影响膀胱癌细胞的增殖进展。结论 LncMALAT1通过竞争性结合miR-506-3p调控EZH2促进膀胱癌增殖功能,进而加快膀胱癌细胞的增殖进展, 可为膀胱癌的治疗提供新的潜在靶点。

Objective To explore the mechanism of long non-coding ribonucleic acid metastasis - associated lung adenocarcinoma transcript 1（LncMALAT1）regulating enhancer of Zeste homolog 2（EZH2）through competitive combination with microRNA-506-3p（miR-506-3p）to affect the proliferation of bladder cancer．Methods A total of 92 pairs of bladder cancer tissues and corresponding adjacent normal tissues were collected from surgical resections between January 2023 and October 2024．The expression levels of LncMALAT1 and EZH2 were detected using Western blot and qRT-PCR．The patients were divided into poor group（n=34）and good group（n=58）according to their prognosis．Clinical data, such as gender, age, tumor diameter, vascular invasion, TNM stage, and distant metastasis were collected, and the relationship between LncMALAT1 and EZH2 and the prognosis of bladder cancer patients was analyzed with clinical pathological indicators．Through in vitro experiments, including qRT-PCR Western blot, plate cloning and EdU experiment were conducted to verify the effect of LncMALAT1 on EZH2 expression and bladder cancer cell proliferation．Bioinformatics technology was used to predict the interaction between LncMALAT1 and miR-506-3p, and qRT-PCR was used to verify the change of miR-506-3p expression after up regulating LncMALAT1 expression in bladder cancer cells．Results The univariate results showed that vascular invasion, TNM stage, distant metastasis, LncMALAT1 and EZH2 expression levels were related to the poor prognosis of bladder cancer patients, and the difference was statistically significant（all P<0.05）．The results showed that the expression of LncMALAT1 and EZH2 in bladder cancer was positively correlated．In vitro experiment results showed that after up regulating LncMALAT1 expression, EZH2 expression was significantly up-regulated, and the proliferation ability of bladder cancer cells was significantly improved（all P<0.05）．QRT-PCR validation showed that the expression of miR-506-3p was significantly down regulated after the expression of LncMALAT1 was up-regulated（P<0.05）, suggesting that LncMALAT1 could regulate EZH2 through competitive combination with miR-506-3p, thereby affecting the proliferation and progression of bladder cancer cells．Conclusions LncMALAT1 can promote the proliferation of bladder cancer cells by competitively combining with miR-506-3p to regulate EZH2, and then accelerate the proliferation of bladder cancer cells, which can provide a new potential target for the treatment of bladder cancer．

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．

目的 检测微小RNA（miR）在人黑色素瘤中的表达情况,研究miR-412通过抑制性别确定区Y框转录因子6（SOX6）的表达影响黑色素瘤细胞增殖及侵袭能力的变化。方法 癌症基因组图谱（TCGA）数据库分析发现miR-412在黑色素瘤中异常表达,为研究其表达与肿瘤的相关性,采用Transwell小室,非锚定独立生长实验分析miR-412对黑色素瘤细胞增殖及侵袭能力的影响。软件预测SOX6可能为其靶向基因,采用荧光素酶报告分析及Western blot实验检测SOX6与miR-412的靶向调节情况。结果 TCGA数据库分析黑色素瘤组织中miR-412表达水平高于正常对照组,表达越高,生存时间越短。Transwell小室,非锚定独立生长实验显示miR-412过表达后促进细胞增殖及侵袭能力,而下调miR-412后抑制黑色素瘤细胞增殖及侵袭能力;通过靶点预测miR-412结合SOX6基因3’-非翻译区（UTR）,导致SOX6蛋白因miR-412表达增高而下调;同时在miR-412下调的细胞中抑制SOX6表达可恢复黑色素瘤细胞的增殖及侵袭能力。结论 miR-412过表达后促进黑色素瘤细胞增殖及侵袭能力,反之则抑制黑色素瘤细胞增殖及侵袭能力。 miR-412通过靶向调控SOX6影响黑色素瘤细胞增殖及侵袭,提示miR-412在黑色素瘤的发病过程中起重要作用,是潜在的治疗靶点。

Objective To assess the expression of miR-412 in human melanoma and investigate how miR-412 modulates melanoma cell proliferation and invasive capacity by inhibiting SRY-Box Transcription Factor 6,（SOX6） expression．Methods Analysis of the TCGA（The Cancer Genome Atlas）database identified aberrant miR-412 expression in melanoma．To explore its relevance to tumorigenesis,we conducted Transwell chamber and non-adherent independent growth assays to examine the effects of miR-412 on melanoma cell proliferation and invasion．Software predictions highlighted SOX6 as a potential target gene．We performed luciferase reporter assays and Western blot experiments to elucidate the regulatory interactions between SOX6 and miR-412．Results TCGA database analysis revealed significantly elevated miR-412 expression levels in melanoma tissues compared to the normal control group．Moreover,higher miR-412 expression correlated with shorter survival times．Functional assays using Transwell chambers and non-adherent independent growth assays demonstrated that overexpressing miR-412 enhanced cell proliferation and invasive capabilities．Conversely,reducing miR-412 expression restrained these attributes in melanoma cells． Target prediction analysis indicated that miR-412 binds to the 3’-UTR region of SOX6,resulting in decreased SOX6 protein levels due to increased miR-412 expression．Intriguingly,inhibiting SOX6 expression concurrently amplified the proliferation and invasive potential of melanoma cells,which was initially dampened by miR-412 downregulation．Conclusions Elevated miR-412 expression augments melanoma cell proliferation and invasive capabilities,while its suppression diminishes these traits．Through its targeted regulation of SOX6,miR-412 exerts a significant influence on melanoma cell proliferation and invasion．These findings underscore the pivotal role of miR-412 in melanoma pathogenesis and underscore its potential as a promising therapeutic target．

目的 探讨微RNA-29b（miR-29b）对子宫内膜癌细胞增殖、迁移和侵袭的影响。方法 子宫内膜癌HEC-1-B细胞分为miR-29b模拟物组（MM组）、miR-29b阻遏物组（MR组）和阴性对照物组（MNC组）,分别转染miR-29b拟似物、miR-29b阻遏物和miR-29b阴性对照物,每组设置6个复孔。以实时定量逆转录PCR检测miR-29b表达,以水溶性四氮唑（WST-1）检测miR-29b对HEC-1-B子宫内膜癌细胞增殖的影响,以Transwell小室检测HEC-1-B子宫内膜癌细胞迁移和侵袭的影响,以Western blot法检测磷酸酶张力蛋白同源物（PTEN）-蛋白激酶 B（AKT）通路蛋白表达水平。结果 MNC组、MM组、MR组miR-29b相对表达量分别为（2 032.1±873.4）、（19 272.8±2 087.9）、（472.7±105.6）,组间比较差异有统计学意义（P<0.05）。MM组0、3、5、7 d时OD值分别为（0.32±0.06）、（0.53±0.08）、（1.13±0.12）和（1.92±0.14）,MNC组0、3、5、7 d时OD值分别为（0.34±0.09）、（0.71±0.08）、（1.67±0.21）和（3.49±0.24）,MR组0、3、5、7 d时OD值分别为（0.38±0.09）、（0.84±0.18）、（2.43±0.24）和（5.67±0.15）,3组0 d时OD值比较差异无统计学意义（P=0.216）,三组3 d、5 d、7 d时OD值比较差异存在统计学意义（P<0.001）。MNC组、MM组和MR组迁移细胞数分别为（403.9±23.8）（102.6±15.7）和（685.7±46.8）个,上述3组侵袭细胞数分别为（82.1±12.7）（38.2±10.6）和（124.6±21.6）个,MM组和MNC组上述指标比较差异均有统计学意义（P<0.05）,MR组和MM组上述指标比较差异均有统计学意义（P<0.05）。MNC组、MM组、MR组PTEN蛋白相对表达量分别为（0.25±0.08）、（0.69±0.11）、（0.11±0.05）,上述3组p-AKT蛋白相对表达量分别为（0.58±0.10）、（0.13±0.06）和（0.79±0.08）,上述3组AKT蛋白相对表达量分别为（0.38±0.09）、（0.37±0.11）和（0.37±0.08）,MM组与MNC组PTEN、p-AKT水平比较差异有统计学意义（P<0.05）,AKT水平比较差异无统计学意义（P>0.05）;MR组与MNC组、MM组PTEN、p-AKT水平比较差异有统计学意义（P<0.05）,AKT水平比较差异无统计学意义（P>0.05）。结论 过表达miR-29b对子宫内膜癌细胞增殖、迁移和侵袭具有抑制作用,靶向PTEN-AKT可能是其重要作用途径。

Objective To investigate the effects of microRNA-29b on proliferation,migration and invasion of endometrial cancer cells．Methods The endometrial cancer HEC-1-B cells were divided into micro29b mimetic group（MM group）,micro29b repressor group（MR group）and negative control group（MNC group）,and the micro29b mimetic,micro29b repressor and micro29b negative control were transfected into each group,six compound holes with each group．The real-time quantitative reverse transcription polymerase chain reaction（RT-PCR）was used to detect the expression of mi29b,WST-1 was used to detect the effect of mi29b on the proliferation of HEC-1-B endometrial cancer cells,Transwell chamber was used to detect the migration and invasion of HEC-1-B endometrial cancer cells,and Western blot was used to detect the expression level of PTEN-AKT pathway protein．Results The relative expression levels of microRNA-29b in MNC group,MM group and MR group were（2 032.1±873.4）,（19 272.8±2 087.9）and（472.7±105.6）,respectively,and there were significant differences between groups（P<0.05）．OD values of MM group at 0 d,3 d,5 d and 7 d were（0.32±0.06）,（0.53±0.08）,（1.13±0.12）and（1.92±0.14）respectively．The OD values of MNC group at 0,3,5 and 7 days were（0.34±0.09）,（0.71±0.08）,（1.67±0.21）and（3.49±0.24）respectively．The OD values of MR group at 0 d,3 d,5 d and 7 d were（0.38±0.09）,（0.84±0.18）,（2.43±0.24）and（5.67±0.15）respectively．There was no significant difference in OD value between the three groups on day 0 （P=0.216）．There were significant differences in OD value between the three groups on day 3,day 5 and day 7（P<0.001）．The number of migrating cells in MNC group,MM group and MR group were（403.9±23.8）cells,（102.6±15.7）cells and（685.7±46.8）cells,respectively．The number of invasive cells in the above three groups were（82.1±12.7）cells,（38.2±10.6）cells and（124.6±21.6）cells．There were significant differences in the above indexes between MM group and MNC group（P<0.05）,also between MR group and MM group（P<0.05）．The relative expression levels of PTEN protein in MNC group,MM group and MR group were（0.25±0.08）,（0.69±0.11）and（0.11±0.05）．The relative expression levels of p-AKT protein in the above three groups were（0.58±0.10）,（0.13±0.06）and（0.79±0.08）．The relative expression levels of AKT protein in the above three groups were（0.38±0.09）,（0.37±0.11）and（0.37±0.08）,respectively．Compared with MNC group,the levels of PTEN and p-AKT in MM group had statistical significance（P<0.05）,but there was no statistical difference in AKT level（P>0.05）．Compared with MNC group and MM group,the levels of PTEN and p-AKT in MR group had statistical significance（P<0.05）,and there was no statistical difference in AKT level（P>0.05）．Conclusions Overexpression of microRNA-29b can inhibit the proliferation,migration and invasion of endometrial cancer cells,and targeting PTEN-AKT may be an important pathway．

目的 探讨柚皮素对人乳腺癌细胞株MCF-7和小鼠乳腺癌细胞系4T1的作用机制。方法 选择人乳腺癌细胞株MCF-7和小鼠乳腺癌细胞系4T1为实验对象,设置对照组和柚皮素组,其中柚皮素组分为20、40、80 和120 μmol/L 4个浓度,利用CCK-8、平板克隆形成实验检测柚皮素对乳腺癌细胞的增殖作用,应用流式细胞术检测柚皮素对乳腺癌细胞的凋亡作用。建立乳腺癌移植瘤模型,应用柚皮素作用于模型小鼠,探讨柚皮素在体内抗肿瘤作用。通过荧光定量PCR和蛋白免疫印迹实验检测自噬相关基因,分析其作用机制。结果 经柚皮素处理后,乳腺癌细胞的增殖明显受到抑制,正常乳腺癌细胞增殖情况变化不大,MCF-7乳腺癌细胞和小鼠乳腺癌4T1均出现明显的凋亡（P<0.001）。结论 柚皮素可以抑制乳腺癌细胞的增殖,且对正常乳腺细胞无明显毒副作用。柚皮素通过凋亡和自噬方式促进乳腺癌细胞的死亡,体内实验结果显示柚皮素具有抗肿瘤作用,并可促进其坏死。

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 体外细胞实验检测吴茱萸碱对骨肉瘤HOS细胞株细胞周期及体外增殖能力的影响。方法 通过利用浓度为0、3、6、12 μmol/L吴茱萸碱处理骨肉瘤HOS细胞48 h后,Hoechst-33258荧光染色观察不同浓度吴茱萸碱处理后HOS细胞核的形态学变化。利用流式细胞术检测3 μmol/L的吴茱萸碱处理后骨肉瘤HOS细胞的细胞周期分布变化。结果 3、6、12 μmol/L的骨肉瘤吴茱萸碱处理细胞,细胞呈现凋亡核碎裂等典型变化,而且随药物剂量增加而更趋明显。呈剂量依赖性抑制其体外增殖能力。3 μmol/L吴茱萸碱处理骨肉瘤HOS细胞0、24、48、72 h,各组细胞周期变化如下:G0/G1期:对照组(51.12±2.13)%、24 h(19.17±1.02)%、48 h(16.94±1.67)%、72 h(11.05±1.25)%;S期:对照组(32.92±0.73)%、24 h(31.00±1.42)%、48 h(32.38±3.03)%、72 h(29.18±2.87)%;G2/M期:对照组(16.01±2.26)%、24 h(49.82±0.62)%、48 h(50.6767±2.80)%、72 h(59.56±1.97)%。结论 吴茱萸碱可诱导人骨肉瘤HOS细胞发生G2/M期阻滞,而S期变化不明显。说明吴茱萸碱可以抑制骨肉瘤细胞的增殖能力,并阻滞细胞周期于G2/M期。

Objective Using transcriptome sequencing and in vitro cell assay to detect the effect of evodiamine on cell cycle and proliferation in osteosarcoma HOS cell line. Methods HOS cells were treated with evodiamine at 0, 3, 6, and 12 μmol/L for 48 hours, Hoechst-33258 fluorescence staining was used to observe the morphological changes of HOS nuclei after treatment with different concentrations of evodiamine.The cell cycle distribution of HOS cells treated with 3 μmol/L evodiamine was detected by flow cytometry. Results 3,6,12 μmol/L osteosarcoma treated with evodiamine, the cells showed typical changes such as apoptotic nuclear fragmentation, and it became more obvious with the increase of drug dosage. Inhibition of proliferation in vitro in a dose-dependent manner.HOS cells were treated with 3 μmol/L evodiamine for 0, 24, 48, 72 h. The cell cycle changes of each group were as follows: G0/G1 phase: control group(51.12±2.13)%, 24 h(19.17±1.02)%, 48 h(16.94±1.67) %, 72 h(11.05±1.25)%;S phase: control group(32.92±0.73)%, 24 h(31.00±1.42)%, 48 h(32.38±3.03)%, 72 h(29.18±2.87)%;G2/M period: control group(16.01±2.26)%, 24 h(49.82±0.62)%, 48 h(50.6767±2.80)%, 72 h(59.56±1.97)%. Conclusion Analysis of the above results revealed that evodiamine can induce G2/M phase arrest in human osteosarcoma HOS cells, but the S phase changes are not obvious. It indicated that evodiamine would inhibit the proliferation of osteosarcoma cells and block the cell cycle in G2/M phase.

目的 研究过表达miR-29a-3p对IL-22诱导的HaCaT细胞增殖的影响。方法 将HaCaT细胞分为Cell组、IL-22组、IL-22+NC组和IL-22+ miR-29a-3p组,荧光定量PCR检测miR-29a-3p的表达水平,CCK8检测细胞的活力,流式细胞仪检测细胞凋亡及周期。结果 与0 μg/L组相比,25 μg/L组、50 μg/L组和100 μg/L组HaCaT细胞的增殖率在24 h、48 h和72 h均出现升高(F值分别为33.27、36.19、52.29,均P<0.000 1)。与0 μg/L组相比,miR-29a-3p在50 μg/L组和100 μg/L组HaCaT中的表达水平降低(F=129,P<0.000 1),分别降低83%和80%。与IL-22+NC组相比,IL-22+ miR-29a-3p组的增殖率在24 h、48 h和72 h均降低(P值分别为0.002 1、0.001 6、0.023 1),细胞总凋亡率增加(6.67±1.06 vs 30.55±1.86,P=0.000 1),G1期细胞比例增加(P=0.000 1),S期细胞比例降低(P=0.000 1)。结论 IL-22可降低HaCaT中miR-29a-3p的表达量,过表达miR-29a-3p通过促进凋亡和引起细胞G1期阻滞抑制IL-22诱发的HaCaT细胞过度增生。

Objective To investigate the effect of miR-29a-3p overexpression on IL-22-induced proliferation of HaCaT cells. Methods HaCaT cells were divided into four groups, Cell group, IL-22 group, IL-22 +NC group and IL-22+miR-29a-3p group. The expression level of miR-29a-3p was detected by fluorescence quantitative PCR. Cell viability was detected by CCK8. Apoptosis and cell cycle were detected by flow cytometry. Results Compared with the 0 g/L group, the proliferation rate of HaCaT cells in the 25 μg/L group, 50 μg/L group and 100 μg/L group was increased at 24 h, 48 h and 72 h (F value was 33.27, 36.19, 52.29,respectively, all P<0.000 1). Compared with the 0 μg/L group, miR-29a-3p expression level in HaCaT in 50 μg/L and 100 μg/L groups was decreased (F=129, P<0.000 1), with a decrease of 83% and 80%, respectively. Compared with the IL-22+NC group, proliferation rate of IL-22+miR-29a-3p group was decreased at 24 h, 48 h and 72 h (P value was 0.002 1, 0.001 6, 0.023 1, respectively), total apoptosis rate was increased (6.67±1.06 vs 30.55±1.86, P=0.000 1), cell proportion in G1 phase was increased (P=0.000 1), and the cell proportion in S phase was decreased (P=0.000 1). Conclusion Il-22 can reduce miR-29a-3p expression level in HaCaT, and miR-29a-3p overexpression can inhibit the excessive proliferation induced by IL-22 in HaCaT cells by promoting apoptosis and inducing G1 phase arrest.

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 本研究从细胞生物学角度检测二甲双胍对小鼠胰岛瘤MIN6的影响,并探讨此过程中包含的分子生物学机制。方法 MTT法检测不同浓度二甲双胍(1、2、5、10、20 mmol/L)对MIN6细胞活力的影响,细胞划痕实验检测二甲双胍对MIN6细胞迁移的影响,免疫印记实验检测此过程中细胞凋亡相关蛋白Bcl-2、Bax、caspase3表达的变化,及AMPK和JNK信号通路蛋白磷酸化水平的变化。结果 二甲双胍浓度大于10 mmol/L时可以抑制MIN6细胞的活力(P<0.01),降低其迁移能力(P<0.01),高浓度二甲双胍可以上调细胞内凋亡蛋白Bax(P<0.05)和p-AMPK的表达(P<0.05),降低抗凋亡蛋白Bcl-2的表达,增加caspase3剪切体(P<0.05)。同时,二甲双胍可以降低MIN6细胞内JNK信号通路的磷酸化水平(P<0.05)。结论 高浓度二甲双胍可以抑制MIN6细胞的增殖和迁移,其作用可能与降低了JNK信号的通路活化有关。

Objective This study aims to investigate the effect of metformin on proliferation and migration of MIN6 cells, and to explore the underlying mechanism. Methods The viability of MIN6 cells that were treated with various metformin (1,2,5,10 and 20 mmol/L) was detected by MTT assay. The migration of MIN6 cells was determined by wound-healing assay. Meanwhile, the proteins expression of Bcl-2, Bax, caspase3 and the phosphorylation of AMPK, JNK was detected by western bolt assay. Results The cell viability and the migration of MIN6 cells were decreased when the concentration of metformin above 10 mmol/L(P<0.01). The expression of apoptosis-related protein Bax(P<0.05) and p-AMPK(P<0.05)was up-regulated, anti-apoptosis-related protein Bcl-2 was down-regulated and cleaved caspase3 (P<0.05)was increased after high metformin treatment. At the same time, the phosphorylation of JNK was down-regulated by metformin(P<0.05). Conclusion High concertration of metformin may inhibit the proliferation and migration of MIN6 cells through suppressing the activation of JNK signaling pathway.