Amentoflavone induces apoptosis and suppresses proliferation of thyroid carcinoma SW579 cells by inhibiting the activation of JAK2-STAT3 pathway / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨阿曼托双黄酮（AF）对甲状腺癌SW579细胞中JAK2-STAT3通路活化及其细胞增殖和凋亡的影响。方法：用0、50、100、150、200 μmol/L的AF处理SW579细胞24、48、72 h，采用CCK-8和Celigo计数、FCM、WB及qPCR法检测AF对SW579细胞的增殖、凋亡、JAK2-STAT3通路活化及其下游调控基因c-Myc、Bcl2、survivin的mRNA及蛋白表达水平的影响。结果：AF处理后，SW579细胞增殖能力显著下降（P<0.05）且呈浓度依赖性，细胞凋亡呈浓度依赖性增多（P<0.05），细胞中JAK2-STAT3通路的活化受到显著抑制（P<0.05），其下游基因c-Myc、Bcl2、survivin的mRNA及蛋白表达均明显下降（均P<0.05）。结论：AF可通过抑制SW579细胞中JAK2-STAT3通路活化及其下游基因的表达而抑制SW579细胞的增殖并促进其凋亡，有望成为治疗甲状腺癌的有效药物。

miRNA-126 regulates the proliferation, apoptosis and migration of thyroid cancer cell SW579 by regulating the notch-1/Akt signaling pathway / 中华内分泌外科杂志

Objective:To investigate the effect of miRNA-126 on proliferation, apoptosis and migration of thyroid cancer SW579 cells, and further to explore its mechanism.Methods:Thyroid cancer SW579 cells were cultured in vitro, and the expression of miRNA-126 in normal thyroid Nthy-ori3-1 cells and SW579 cells was detected by qPCR. The cells were divided into blank control group (without any treatment) , experimental group (transfected with miRNA-126 expression plasmid) and negative control group (transfected with empty plasmid vector) . SW579 cells with overexpression of miRNA-126 were constructed by transfection of plasmids. CCK-8 test was used to detect cell proliferation; Transwell cell test was used to detect cell migration and invasion; Flow cytometry was employed to detect apoptosis and changes in reactive oxygen levels; Western blot was used to detect Notch-1/Akt pathway related protein expression.Results:The expression level of miRNA-126 in SW579 cells was 0.25±0.07, and the difference was statistically significant compared with that in Nthy-ori3-1 cells (P<0.001) . The cell survival rates of blank control group, experimental group and negative control group were (105.70 ± 7.61) , (98.60 ± 5.42) and (62.70 ± 3.82) ; The apoptosis rates were (9.14 ± 0.83) , (12.28 ± 1.34) and (36.39 ± 3.21) (all P < 0.05) ; The cell migration rates were (34.51 ± 2.45) , (33.29 ± 3.17) and (11.22 ± 1.23) (all P < 0.05) ; The levels of ROS were (1.02 ± 0.07) , (1.08 ± 0.11) and (6.54 ± 0.74) (all P < 0.05) .Conclusion:Overexpression of miR-NA-126 can inhibit Notch-1/Akt pathway by up-regulating intracellular reactive oxygen species, inhibit the proliferation and migration of SW579 cells, and induce apoptosis in SW579 cells, which provides a certain basis for the study of miRNA-126 in thyroid cancer.

lncRNA SNHG14 regulates the malignant biological behaviors of thyroid cancer SW579 cells by targeting miR-433-3p / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨lncRNA SNHG14对甲状腺癌SW579细胞恶性生物学行为的影响及其分子机制。方法：收集2017年10月至2018年12月青海省人民医院收治的20例甲状腺癌患者的癌组织及癌旁组织标本，用qPCR检测甲状腺癌组织和对应癌旁组织中SNHG14与miR-433-3p的表达；根据转染物的不同，将SW579细胞分为si-NC组（转染si-NC）、si-SNHG14组（转染si-SNHG14）、miR-NC组（转染miR-NC）、miR-433-3p mimic组（转染miR-433-3p mimic）、si-SNHG14+anti-miR-NC组（共转染si-SNHG14与anti-miR-NC）和si-SNHG14+anti-miR-433-3p组（共转染si-SNHG14与anti-miR-433-3p）。MTT法、FCM、Transwell实验分别检测转染后SW579细胞的增殖能力、细胞周期、细胞凋亡率、迁移及侵袭能力的改变；利用双荧光素酶报告基因实验检测SNHG14是否可结合miR-433-3p，qPCR法检测SNHG14与miR-433-3p之间的相互调控关系。结果：SNHG14在甲状腺癌组织中的表达高于癌旁组织（P<0.05），而miR-433-3p的表达水平低于癌旁组织（P<0.05）。抑制SNHG14的表达或过表达miR-433-3p可使SW579细胞增殖能力降低（P<0.05）、迁移与侵袭细胞数减少（均P<0.05）、细胞凋亡率升高（P<0.05）、G1期细胞比例升高（P<0.05）且S期细胞比例降低（P<0.05）。双荧光素酶报告基因实验证明SNHG14可结合miR-433-3p，抑制SNHG14的表达可提高SW579细胞中miR-433-3p水平（均P<0.05）。同时抑制miR-433-3p 和SNHG14的表达可部分逆转后者对SW579细胞的增殖、凋亡、迁移和侵袭的作用（均P<0.05）。结论：甲状腺癌组织中lncRNA SNHG14呈高表达、miR-433-3p呈低表达，lncRNA SNHG14可通过靶向结合miR-433-3p促进甲状腺癌SW579细胞的增殖、迁移、侵袭而抑制细胞凋亡。

Iodine-131 treatment of thyroid cancer cells leads to suppression of cell proliferation followed by induction of cell apoptosis and cell cycle arrest by regulation of B-cell translocation gene 2-mediated JNK/NF-kappaB pathways

Iodine-131 (131I) is widely used for the treatment of thyroid-related diseases. This study aimed to investigate the expression of p53 and BTG2 genes following 131I therapy in thyroid cancer cell line SW579 and the possible underlying mechanism. SW579 human thyroid squamous carcinoma cells were cultured and treated with 131I. They were then assessed for 131I uptake, cell viability, apoptosis, cell cycle arrest, p53 expression, and BTG2 gene expression. SW579 cells were transfected with BTG2 siRNA, p53 siRNA and siNC and were then examined for the same aforementioned parameters. When treated with a JNK inhibitor of SP600125 and 131I or with a NF-κB inhibitor of BMS-345541 and 131I, non-transfected SW579 cells were assessed in JNK/NFκB pathways. It was observed that 131I significantly inhibited cell proliferation, promoted cell apoptosis and cell cycle arrest. Both BTG2 and p53 expression were enhanced in a dose-dependent manner. An increase in cell viability by up-regulation in Bcl2 gene, a decrease in apoptosis by enhanced CDK2 gene expression and a decrease in cell cycle arrest at G0/G1 phase were also observed in SW579 cell lines transfected with silenced BTG2 gene. When treated with SP600125 and 131I, the non-transfected SW579 cell lines significantly inhibited JNK pathway, NF-κB pathway and the expression of BTG2. However, when treated with BMS-345541 and 131I, only the NF-κB pathway was suppressed. 131I suppressed cell proliferation, induced cell apoptosis, and promoted cell cycle arrest of thyroid cancer cells by up-regulating B-cell translocation gene 2-mediated activation of JNK/NF-κB pathways.

Effects of downregulation of CD147 in the mobility of thyroid cancer cell SW579 and its mechanisms / 实用医学杂志

Objective To detect the effects and mechanisms of CD147 in thyroid cancer cell SW579. Methods RT-PCR and Western-blot were used to determine the levels of CD147 mRNA and protein in SW579 cells after treated with CD147 specific small interference RNA (siRNA). The proliferative ratio of SW579 cells after treated with CD147 siRNA was detected by CCK-8 assay. We detected the changes of cellular mobility after treated with CD147 siRNA by transwell assay. Mobility related proteins were analyzed by western-blot. Results The proliferative ratio and the mobility of SW579 cells were inhibited significantly by down-regulation of CD147. The levels of MMP-2 and MMP-9 were lower in treated cells than untreated ones. Expression of Wave 2 and a mobility related protein were decreased in accompany with down-regulation of CD147. Conclusion Down-regulation of CD147 inhibits mobility of SW579 by suppressing MMP-2, MMP-9 and WAVE 2.

Effects of Paclitaxel Combined with Cisplatin on the Proliferation,Migration and Invasion of Thyroid Can-cer Cells SW579 / 中国药房

OBJECTIVE:To study the effects of paclitaxel combined with cisplatin on the proliferation,migration and invasion of thyroid cancer cells SW579 and its mechanism. METHODS:Cells were divided into blank control group,paclitaxel group (3μmol/L),cisplatin group(30 μmol/L),drug combination group(paclitaxel 3 μmol/L+cisplatin 30 μmol/L). 48 h after culture,the relative cell activity was measured by MTT assay. Cell cycle was detected by flow cytometry. Migration and invasion of cell was tested by Transwell assay. The expression of phosphatase and tensin homolog deleted on chromosome ten(PTEN),protein kinase B(AKT),Cyclin D1,p27,matrix metalloproteinase(MMP)-2 and MMP-9 were detected by Western blot. RESULTS:Compared with blank control group,relative cell activity of all treatment groups were decreased;paclitaxel or plus cisplatin also made cell cy-cle arrest in G1 phase,and migration and invasion ability of cell were decreased;the expression of PTEN and p27 remarkably in-creased,while the expression of Cyclin D1,MMP-2,MMP-9 and phosphorylation of AKT were obviously reduced,with statisti-cal significance (P<0.05). Compared with single drug group,the effect of drug combination group strengthened,with statistical significance in above indicators(P<0.05). CONCLUSIONS:The inhibition effect of paclitaxel combined with cisplatin on the pro-liferation,migration and invasion of thyroid cancer cells SW579 cell will be strengthened,by a mechanism of up-regulating the ex-pression of PTEN and p27,down-regulating the expression of Cyclin D1,MMP-2 and MMP-9,inhibiting phosphorylation of AKT.

REGγ is a strong candidate for the regulation of cell cycle, proliferation and the invasion by poorly differentiated thyroid carcinoma cells

REGγ is a proteasome activator that facilitates the degradation of small peptides. Abnormally high expression of REGγ has been observed in thyroid carcinomas. The purpose of the present study was to explore the role of REGγ in poorly differentiated thyroid carcinoma (PDTC). For this purpose, small interfering RNA (siRNA) was introduced to down-regulate the level of REGγ in the PDTC cell line SW579. Down-regulation of REGγ at the mRNA and protein levels was confirmed by RT-PCR and Western blot analyses. FACS analysis revealed cell cycle arrest at the G1/S transition, the MTT assay showed inhibition of cell proliferation, and the Transwell assay showed restricted cell invasion. Furthermore, the expression of the p21 protein was increased, the expression of proliferating cell nuclear antigen (PCNA) protein decreased, and the expression of the p27 protein was unchanged as shown by Western blot analyses. REGγ plays a critical role in the cell cycle, proliferation and invasion of SW579 cells. The alteration of p21 and PCNA proteins related to the down-regulation of REGγ suggests that p21 and PCNA participate in the process of REGγ regulation of cell cycle progression and cell proliferation. Thus, targeting REGγ has a therapeutic potential in the management of PDTC patients.