姜黄素增强人乳头瘤状甲状腺癌细胞TPC-1放射敏感性的研究

目的 研究姜黄素对人乳头瘤状甲状腺癌细胞TCP-1放射敏感性的影响,并探索姜黄素可能作用的信号通路,为甲状腺癌放射增敏剂的开发提供新的思路。方法 使用姜黄素和放射性碘处理人乳头瘤状甲状腺癌细胞TPC-1,CCK-8法检测细胞增殖,克隆形成实验检测细胞克隆形成能力,流式细胞仪检测细胞凋亡情况,western blot检测p50、p65、凋亡相关蛋白Bcl-2和Bax的表达变化。使用NF-κB信号通路抑制剂PDTC抑制NF-κB信号通路活性,检测细胞增殖、克隆形成和凋亡变化。结果 姜黄素和放射性碘处理人乳头瘤状甲状腺癌细胞TPC-1后,细胞增殖率下降,克隆形成减少,凋亡上升,凋亡抑制蛋白Bcl-2表达下调,促凋亡蛋白Bax表达上调,并呈浓度依赖性。这些结果表明姜黄素可增加TPC-1细胞的放射敏感性。碘放射后TPC-1细胞NF-κB通路活化,姜黄素可以抑制碘放射后TPC-1细胞NF-κB通路的激活。使用抑制剂抑制NF-κB通路的活性,细胞增殖下降,克隆形成减少,凋亡上升,放射敏感性升高。结论 姜黄素可能靶向NF-κB信号通路,通过抑制NF-κB信号通路活性调节甲状腺癌细胞的放射敏感性。     

Polycomb-mediated loss of miR-31 activates NIK-dependent NF-κB pathway in adult T cell leukemia and other cancers

Constitutive NF-κB activation has causative roles in adult T cell leukemia (ATL) caused by HTLV-1 and other cancers. Here, we report a pathway involving Polycomb-mediated miRNA silencing and NF-κB activation. We determine the miRNA signatures and reveal miR-31 loss in primary ATL cells. MiR-31 negatively regulates the noncanonical NF-κB pathway by targeting NF-κB inducing kinase (NIK). Loss of miR-31 therefore triggers oncogenic signaling. In ATL cells, miR-31 level is epigenetically regulated, and aberrant upregulation of Polycomb proteins contribute to miR-31 downregulation in an epigenetic fashion, leading to activation of NF-κB and apoptosis resistance. Furthermore, this emerging circuit operates in other cancers and receptor-initiated NF-κB cascade. Our findings provide a perspective involving the epigenetic program, inflammatory responses, and oncogenic signaling.     

Rab11通过NF-κB信号通路调控膀胱癌细胞的增殖和侵袭

目的:探讨Rab11调控膀胱癌细胞增殖和侵袭的分子机制。方法:用Western blot方法检测膀胱癌BIU-87、T24、RT4、5637细胞系中Rab11的内源表达量。在BIU-87（Rab11低表达）和T24（Rab11高表达）细胞系中分别转染Rab11过表达质粒和Rab11 siRNA。用荧光素酶报告实验检测转染后细胞内的NF-κB信号通路的活性,通过Western blot技术分析与NF-κB信号通路相关蛋白p-IκB的变化情况。用NF-κB抑制剂Bay 11-7082处理转染Rab11过表达质粒的BIU-87细胞系,通过Western blot检测细胞系中细胞周期相关因子cyclin D1和侵袭相关因子MMP9蛋白的变化。结果:转染Rab11过表达质粒的BIU-87细胞系中NF-κB活性水平提高,p-IκB表达量显著提高;而敲除Rab11后NF-κB活性受到抑制,p-IκB的表达受到抑制,而p-IκB与NF-κB信号通路的活性呈正相关。NF-κB抑制剂Bay 11-7082阻遏了Rab11诱导cyclin D1和MMP9表达上调的过程。结论:Rab11通过NF-κB信号通路调控膀胱癌细胞的增殖和侵袭。     

miR-224-5p靶向调控KIF23通过NF-κB信号通路影响宫颈癌细胞增殖、迁移和侵袭

目的:探索微小RNA-224-5p(miR-224-5p)对驱动蛋白超家族23(KIF23)的靶向关系及对宫颈癌细胞增殖、迁移和侵袭的影响。方法:采用实时定量PCR(qPCR)检测正常宫颈细胞(H8)和宫颈癌细胞(SiHa、HeLa、MS751、HT-3)中miR-224-5p和KIF23 mRNA表达,选择miR-224-5p表达量最低的HeLa细胞开展后续研究。在HeLa细胞中转染si-KIF23或miR-224-5p,探讨敲减KIF23或过表达miR-224-5p对HeLa细胞增殖、迁移和侵袭的影响。噻唑蓝(MTT)比色法检测细胞增殖,Transwell小室法检测细胞迁移和侵袭,蛋白质印迹法(Western Blot)检测KIF23、细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)和NF-κBp65蛋白水平;生物学信息预测和双荧光素酶报告基因实验分析miR-224-5p和KIF23之间的靶向关系;共转染si-KIF23和anti-miR-224-5p,观察miR-224-5p低表达对敲减KIF23诱导的HeLa细胞增殖、迁移、侵袭和NF-κB信号通路活化的影响。结果:miR-224-5p在SiHa、HeLa、MS751、HT-3细胞中表达下调(P<0.05),KIF23 mRNA和蛋白表达上调(P<0.05)。敲减KIF23或过表达miR-224-5p显著降低HeLa细胞存活率、细胞迁移数和侵袭数,并显著抑制CyclinD1、MMP2、MMP9蛋白水平(P<0.05)。另外,敲减KIF23显著降低细胞中NF-κBp65表达量(P<0.05)。miR-224-5p靶向调控KIF23表达。miR-224-5p低表达可以逆转敲减KIF23抑制HeLa增殖、迁移、侵袭及CyclinD1、MMP2、MMP9蛋白表达的作用,以及逆转敲减KIF23抑制NF-κB信号通路活化的作用。结论:miR-224-5p靶向调控KIF23并通过NF-κB信号通路抑制宫颈癌细胞增殖、迁移和侵袭。     

敲减lncRNA RP1-261G23.7抑制胶质瘤细胞增殖和迁移及其机制探讨北大核心

目的:探讨长链非编码RNA(long non-coding RNA,lncRNA)RP1-261G23.7对人胶质瘤细胞增殖及迁移的影响及可能的调控机制。方法:采用GEPIA数据库分析RP1-261G23.7在胶质瘤组织中的相对表达。采用qRT-PCR检测RP1-261G23.7在4种胶质瘤细胞系(U87MG、SNB-19、U251、LN382)中的相对表达。采用Lipofectamine3000向胶质瘤细胞U87MG中单独转染si-RP1-261G23.7质粒(si-RP1-261G23.7组)和si-NC对照质粒(si-NC组)。采用CCK-8实验和细胞划痕实验检测转染后U87MG细胞的增殖及迁移能力。采用生物信息学、qRT-PCR和双荧光素酶报告基因实验研究RP1-261G23.7和miR-525-5p表达的关系。Western blotting检测NF-κB信号通路蛋白的表达。结果:与正常组织相比,RP1-261G23.7在胶质瘤组织中表达上调(P<0.01)。与人脑星型胶质正常细胞系(HEB)相比,RP1-261G23.7在四种胶质瘤细胞系中表达均上调(P<0.01),U87MG细胞中RP1-261G23.7相对表达量最高(P<0.01)。与si-NC组相比,敲减RP1-261G23.7显著抑制了U87MG细胞的增殖(P<0.05)和划痕愈合(P<0.01)。RP1-261G23.7能够直接互补结合miR-525-5p(P<0.01)。与si-NC组相比,敲减RP1-261G23.7表达显著促进了U87MG细胞中miR-525-5p的表达(P<0.01),NF-κB信号通路蛋白表达显著下降(P<0.01)。结论:胶质瘤组织和细胞系中RP1-261G23.7表达明显上调,敲减RP1-261G23.7通过促进miR-525-5p表达、干扰NF-κB信号通路活化,抑制胶质瘤U87MG细胞的增殖和迁移,可能为胶质瘤的靶向治疗开辟新的路径。     

miR-485-3p通过靶向TLR1/NF-κB信号通路调节胃癌细胞放射敏感性

目的 研究miR-485-3p是否通过靶向TLR1对胃癌细胞放射敏感性起作用。方法分别用qRT-PCR和蛋白印迹法检测miR-485-3p和TLR1的表达变化,DIANA、TargetScan和miRanda软件预测和双荧光素酶报告实验验证miR-485-3p对TLR1的靶向作用。将miR-485-3p mimic或TLR1 siRNA转染到胃癌MGC803细胞中,放射处理细胞,凋亡实验、克隆形成实验和MTT检测细胞放射敏感性的变化。双荧光素酶报告实验检测miR-485-3p上调和TLR1沉默对NF-κB活性的影响。蛋白免疫印迹实验探究miR-485-3p上调和TLR1沉默对NF-κB靶基因的影响。结果 放射处理后胃癌细胞miR-485-3p表达下调,TLR1表达上调。靶基因预测软件发现TLR1可能是miR-485-3p的靶基因,双荧光素酶报告实验进一步验证了TLR1是miR-485-3p的直接靶点。miR-485-3p负调控TLR1的表达。miR-485-3p的过表达提高了细胞凋亡率,降低了细胞克隆形成和细胞群体增殖能力,增强了细胞的放射敏感性,且TLR1的沉默也具有相同作用。miR-485-3p上调和TLR1沉默均降低了NF-κB的活性,下调了NF-κB多个靶基因的表达。结论 miR-485-3p可能通过靶向TLR1调控NF-κB信号通路增强胃癌细胞的放射敏感性。     

Sulforaphane Inhibits the Proliferation of the BIU87 Bladder Cancer Cell Line via IGFBP-3 Elevation

Aim: To investigate effects of sulforaphane on the BIU87 cell line and underlying mechanisms involvingIGFBP-3. Methods: Both BIU87 and IGFBP-3-silenced BIU87 cells were treated with sulforaphane. Cellproliferation was detected by MTT assay. Cell cycle and apoptosis were determined via flow cytometry.Quantitative polymerase chain reaction and Western blotting were applied to analyze the expression of IGFBP-3and NF-κB at both mRNA and protein levels. Results: Sulforaphane (80 μM) treatment could inhibit cellproliferation, inducing apoptosis and cell cycle arrest at G2/M phase. All these effects could be antagonized byIGFBP-3 silencing. Furthermore, sulforaphane (80 μM) could down-regulate NF-κB expression while elevatingthat of IGFBP-3. Conclusions: Sulforaphane could suppress the proliferation of BIU87 cells via enhancingIGFBP-3 expression, which negatively regulating the NF-κB signaling pathway.     

Rosmarinic Acid Induces Proliferation Suppression of Hepatoma Cells Associated with NF-κB Signaling Pathway

Background: Rosmarinic acid (RA) is a natural phenolic compound that acts as a Fyn inhibitor by 53 homology modeling of the human Fyn structure. Therefore, the apoptosis mechanism related to  NF-κB signaling pathway induced by RA in HepG2 was investigated. Methods: The cell growth, apoptosis, and proliferation of HepG2 regulated by various concentrations of RA were studied. The proteins expression of MMP-2, MMP-9, PI3K, AKT, NF-κB, and apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3 were detected. Results: RA significantly reduced proliferation rates, inhibited migration and invasion, and decreased the expressions of invasion-related factors, such as matrix metalloproteinase (MMP)-2 and MMP-9. TUNEL staining revealed that RA resulted in a dose-dependent increase of HepG2 cell apoptosis. In line with this finding, the expression of apoptosis suppressor protein Bcl-2 was downregulated and that of the pro-apoptotic proteins Bax and cleaved caspase-3 was increased. In addition, we found that the phosphatidylinositol 3-kinase (PI3K)/Akt/nuclear factor kappa B (NF-κB) signaling pathway was involved in RA-mediated inhibition of HepG2 cell metastasis. Conclusion: Our study identified that  RA as a drug candidate for the treatment of HCC.     

TAK-242对人多发性骨髓瘤细胞增殖及凋亡的影响

 目的 探讨Toll样受体4（TLR4）特异性抑制剂TAK-242对人多发性骨髓瘤细胞株RPMI8226增殖及凋亡的影响。方法 取对数期生长的人多发性骨髓瘤细胞株RPMI8226分为A、B、C组及对照组（Control），A、B、C组分别加入终浓度为20、40、80 μmol/L的TAK-242，对照组不加抑制剂，培养24 h后，采用CCK-8法检测细胞增殖抑制率；Annexin-V/PI检测细胞凋亡率；RT-PCR检测细胞TLR4、Myd88 mRNA的表达水平；Western blot检测细胞Myd88、NF-κB的蛋白表达情况。结果 A、B、C组细胞增殖抑制率、凋亡率升高，TLR4、Myd88 mRNA表达水平及Myd88、NF-κB的蛋白表达水平降低，各浓度组之间的差异均有统计学意义（P<0.05）。结论 TAK-242可抑制人多发性骨髓瘤细胞RPMI8226的增殖，促进细胞凋亡，其机制可能为NF-κB信号通路功能被抑制。     

NGX6基因联用5-Fu对人结肠癌细胞凋亡的影响

目的:探讨候选抑瘤基因NGX6联用5-Fu对结肠癌细胞凋亡的影响.方法:以稳定转染并表达NGX6基因的HT-29细胞与5-Fu联用作为实验组.以PDTC与5-Fu联用的HT-29细胞作为对照组.通过EMSA检测各组结肠癌HT-29细胞核转录因子-κB(NF-κB)的激活情况,利用MTT比色法检测各组细胞增殖的情况.吖啶橙(AO)/溴化乙啶(EB)双染法显微镜观测以及PI/Annexin-V双染流式细胞仪检测各组细胞凋亡情况.结果:稳定转染并表达NGX6基因的HT-29细胞以及应用了PDTC的HT-29细胞NF-κB的激活均明显受到抑制;与5-Fu作用的HT-29细胞组比较,5-Fu联合PDTC作用于HT-29细胞后,HT-29细胞增殖受到明显抑制,5-Fu诱导HT-29细胞凋亡作用增强;与5-Fu联合PDTC作用于HT-29细胞的对照组比较,在诱导细胞凋亡以及抑制细胞增殖方面,稳定转染并表达NGX6基因的HT-29细胞与5-Fu联用组和对照组所取得一致的效果,NGX6基因增强5-Fu对HT-29细胞增殖抑制的能力及诱导HT-29细胞凋亡的能力.结论:NGX6基因抑制了肿瘤细胞NF-κB的激活,具有增强5-Fu诱导结肠癌细胞凋亡的能力,其机制可能是抑制肿瘤细胞NF-κB的激活,NGX6基因对肿瘤的治疗及预后起积极作用.     

IL-12 Regulates B7-H1 Expression in Ovarian Cancerassociated Macrophages by Effects on NF-κB Signalling

Background and Aim: B7-H1, a co-inhibitory molecule of the B7 family, is found aberrantly expressed inovarian cancer cells and infiltrating macrophage/dendritic-like cells, and plays a critical role in immune evasionby ovarian cancer. IL-12, an inducer of Th1 cell development, exerts immunomodulatory effects on ovariancancer. However, whether IL-12 regulates B7-H1 expression in human ovarian cancer associated-macrophageshas not been clarified. Therefore, we investigated the effects of IL-12 on the expression of B7-H1 in ovariancancer-associated macrophages and possible mechanisms. Methods: PMA induced THP-1-derived macrophagesor human monocyte-derived macrophages were treated with recombinant IL-12 (rIL-12) or infected withadenovirus carrying human IL-12 gene (Ad-IL-12-GFP) for 24 h, then cocultured with the SKOV3 ovariancancer cell line for another 24 h. Macrophages were collected for real-time PCR and Western blot to detect theexpression of B7-H1, and activation of the NF-κB signaling pathway. Moreover, supernatants were collected toassay for IL-12, IFN-γ and IL-10 by ELISA. In addition, monocyte-derived macrophages treated with IFN-γ werecocultured with SKOV3 and determined for the expression of B7-H1. Furthermore, the expression of B7-H1 inmonocyte-derived macrophages was also evaluated after blocking NF-κB signaling. Results: The expression ofB7-H1 was significantly upregulated in monocyte-derived macrophages treated with rIL-12 or Ad-IL-12-GFPcompared with the control groups (p<0.05), accompanied by a remarkable upregulation of IFN-γ (p<0.05), amarked downregulation of IL-10 (p<0.05) and activation of NF-κB signaling. However, the upregulation of B7-H1 was inhibited by blocking the NF-κB signaling pathway (p<0.05). Expression of B7-H1 was also increased(p<0.05) in monocyte-derived macrophages treated with IFN-γ and cocultured with SKOV3. By contrast, theexpression of B7-H1 in THP-1-derived macrophages was significantly decreased when treated in the same wayas monocyte-derived macrophages (p<0.05), and IL-10 was also significantly decreased but IFN-γ was almostabsent. Conclusions: IL-12 upregulates the expression of B7-H1 in monocyte-derived macrophages, which ispossible though inducing the secretion of IFN-γ and further activating the NF-κB signal pathway. However,IL-12 downregulates the expression of B7-H1 in THP-1-derived macrophages, associated with a lack of IFN-γand inhibition of expression of IL-10.     

MiR-125b靶向A20/NF-κB信号通路促进胰腺癌细胞增殖机制的研究

[目的]研究miR-125b在胰腺癌组织及细胞系中的表达,探讨其对胰腺癌细胞增殖的影响及作用机制。[方法]QRT-PCR检测miR-125b在60例胰腺癌患者癌组织和癌旁组织中,及3株胰腺癌细胞系和1株正常胰腺导管上皮细胞中的表达;miR-125b敲减慢病毒质粒及阴性对照空载体质粒vector感染SUIT-2细胞,经1.0μg/ml嘌呤霉素筛选稳定敲减miR-125b或vector的SUIT-2细胞,注射到BALB/c裸鼠中,观察抑制miR-125b对胰腺癌细胞体内增殖能力的影响;利用Targetscan 7.2软件及荧光素酶报告基因试验,分析miR-125b对A20基因的靶向作用;SUIT-2细胞转染miR-125b inhibit和scramble后,体外实验检测miR-125b对细胞增殖、肿瘤坏死因子α诱导的蛋白3(tumor necrosis factorα-induced protein 3,TNFAIP3,又称A20)和NF-κB信号通路关键蛋白IKKα/β、IκBα、p65的表达影响。[结果]MiR-125b在胰腺癌的组织中的表达显著高于癌旁组织(0.99±0.21 vs 0.68±0.17,P<0.05);miR-125b在SW1990、SUIT-2、BxPC3胰腺癌细胞系中的表达显著高于在正常胰腺导管上皮细胞HPDE6-C7中的表达(4.81±0.13,8.63±0.27,5.64±0.21 vs1.00±0.05,P均<0.05);SUIT-2_(mi R-125b)的裸鼠成瘤体积显著低于SUIT-2_vector细胞(P<0.05)。TargetScan软件预测和荧光素酶报告基因实验验证A20为miR-125b的靶基因;转染miR-125b inhibit后,A20基因表达显著增加(6.98±0.18 vs 1.00±0.06,P<0.05),细胞增殖能力、NF-κB信号通路关键蛋白IK-Kα/β、IκBα、p65的表达显著下降(P均<0.05)。[结论]MiR-125b可通过靶向调控A20/NF-κB信号通路促进胰腺癌细胞的增殖。     

miR-152在急性髓系白血病患者骨髓和细胞系中的表达及生物学功能的初步研究

目的:检测miR-152在急性髓系白血病(AML)患者骨髓和细胞系中的表达水平。初步研究miR-152在急性髓系白血病中的生物学功能。方法:收集急性髓系白血病患者40例,非恶性血液病对照组20例,提取骨髓有核细胞;培养U937、Kasumi-1、THP-1三种急性髓系白血病细胞系。RT-PCR检测miR-152的表达水平。分别对U937细胞系和Kasumi-1细胞系转染miR-152 mimics和inhibitor,CCK-8法检测U937细胞及Kasumi-1细胞增殖情况,Annexin V/PI流式实验检测细胞凋亡,流式细胞术检测细胞周期。结果:miR-152在急性髓系白血病患者骨髓及细胞系中较非恶性血液病对照组表达明显下降。U937细胞系转染miR-152 mimics后,增殖受抑制,细胞凋亡增加,细胞阻滞在G0-G1期;Kasumi-1细胞系转染miR-152 inhibitor后,增殖增加,细胞凋亡减少,G0-G1期细胞减少。结论:miR-152在急性髓系白血病患者骨髓及细胞系中的表达水平均较正常对照组显著降低。miR-152在急性髓系白血病中起抑癌基因的作用,过表达miR-152可以抑制细胞增殖,促进凋亡。     

MicroRNA-9 suppresses uveal melanoma cell migration and invasion through the NF-κB1 pathway

The aggressive course of uveal melanoma is believed to reflect its unusually invasive and metastatic nature, which is associated with the nuclear factor kappaB (NF-κB) pathway. MicroRNAs (miRNAs) have been implicated in the regulation of various biological and pathological processes in cancer, however, the special role of miR-9 in uveal melanoma metastasis is largely unknown. In the present study, we showed that miR-9 is significantly reduced in highly invasive uveal melanoma cell lines, and suppressed migration and invasion of highly invasive cells. Furthermore, miR-9 negatively modulated NF-κB1 expression by direct targeting at its 3'-UTRs. Additionally, downstream targets of NF-κB1, such as MMP-2, MMP-9 and VEGFA, were regulated by miR-9 in the same pattern as NF-κB1. Therefore, miR-9 suppresses uveal melanoma cell migration and invasion partly through downregulation of the NF-κB1 signaling pathway.     

MicroRNA-653靶向调控OIP5基因介导mTOR信号通路对非小细胞肺癌细胞生物学特性的影响

目的:探讨microRNA-653（miR-653）靶向调控OIP5基因介导mTOR信号通路对非小细胞肺癌（non-small cell lung cancer,NSCLC）细胞生物学特性的影响。方法:选取人正常内皮细胞BEAS-2B和4种NSCLC细胞系（H1650、H1975、A549和H292）,分为control组、mimics组、mimics-NC组、inhibitor组和inhibitor-NC组进行瞬时转染,利用荧光定量PCR、蛋白印迹、MTT、划痕实验、Transwell侵袭实验、流式细胞术等方法分析miR-653对NSCLC细胞中OIP5、mTOR信号通路相关基因表达及增殖、迁移、侵袭、凋亡和细胞周期分布的影响。结果:与control组、mimics-NC组和inhibitor-NC组相比,mimics组mTOR信号通路被抑制,细胞增殖、迁移、侵袭能力明显降低,凋亡率明显增加,细胞多停滞于G1期（均P＜0.05）;然而,inhibitor组mTOR信号通路被激活,细胞增殖、迁移、侵袭能力明显升高,而凋亡率明显降低,G1期细胞数量较少（均P＜0.05）。结论:miR-653可通过负调控OIP5基因抑制mTOR通路激活,从而阻止NSCLC的发生与发展。     

SGPL1在神经胶质瘤组织中的表达及其对细胞增殖和凋亡的影响

目的:探讨SGPL1在神经胶质瘤组织中的表达及其对神经胶质瘤细胞增殖和凋亡的影响。方法:在TCGA及PROGgene在线数据库中分析SGPL1在神经胶质瘤临床肿瘤组织及正常神经组织中的表达差异,并分析其表达与神经胶质瘤预后生存的关系;在神经胶质瘤细胞系U251和U87中分别敲降和过表达SGPL1,通过RT-qPCR和Western blot实验分别检测SGPL1的mRNA和蛋白表达水平;用CCK-8法、细胞克隆形成实验检测SGPL1敲降和过表达后细胞增殖变化;通过流式细胞术和caspase3/7酶活性试剂检测分析细胞凋亡变化,Western blot实验检测caspase3、PARP及CyclinD1等凋亡相关蛋白的表达;ELISA检测细胞内S1P含量;RT-qPCR检测S1PR5的表达水平;RNA-seq分析SGPL1在神经胶质瘤细胞中调控的信号通路;Western blot实验检测SGPL1敲降和过表达后Phospho-p38-MAPK、Phospho-ERK、Phospho-STAT3及Phospho-NF-κB(p65)的表达变化。结果:TCGA及PROGgene在线数据库分析表明SGPL1在神经胶质瘤组织中高表达并与神经胶质瘤患者的预后生存成负相关。在U251中敲降SGPL1后细胞增殖受到抑制,细胞凋亡增加;在U87细胞中过表达SGPL1后能显著促进细胞的增殖能力。SGPL1表达对S1P信号通路无明显影响;RNA-seq结果表明SGPL1在神经胶质瘤细胞内调控细胞增殖死亡信号通路,SGPL1敲降和过表达可影响p38-MAPK、ERK、STAT3及NF-κB(p65)等细胞增殖生长信号的变化。结论:SGPL1在神经胶质瘤组织中高表达并与生存预后负相关。在神经胶质瘤细胞内SGPL1可调控p38-MAPK、ERK、STAT3及NF-κB(p65)等细胞增殖生长信号通路,其表达可影响神经胶质瘤细胞的增殖和凋亡,可能是一个潜在的肿瘤标志物和治疗的分子靶点。     

吴茱萸碱逆转人肺癌细胞株Ａ５４９／ＤＤＰ耐药机理的实验研究

目的　观察吴茱萸碱逆转人肺腺癌耐药株Ａ５４９／ＤＤＰ细胞耐药性的效果并探讨其与阻断ＮＦ κＢ信号传导通路的相关性。方法　采用ＭＴＴ法检测单用吴茱萸碱、顺铂（ＤＤＰ）以及两药联用在不同时间对Ａ５４９／ＤＤＰ细胞的增殖影响,计算ＩＣ５０及耐药逆转倍数。流式细胞仪检测各组细胞的凋亡情况。ＲＴ ＰＣＲ检测各组ＭＤＲ１、ＮＦ κＢ、Ｂｃｌ-２、ＭＭＰ-２和ＶＥＧＦ的ｍＲＮＡ表达。Ｗｅｓｔｅｒｎｂｌｏｔ法检测各组细胞的ｐＩκＢ-α、pＩＫＫα蛋白表达水平。结果　吴茱萸碱０.１２５ｍｇ／Ｌ、０.２５ｍｇ／Ｌ针对Ａ５４９／ＤＤＰ细胞对ＤＤＰ的耐药逆转倍数分别为３.６６８和１1.４８。ＲＴ ＰＣＲ显示在吴茱萸碱作用下检测基因的ｍＲＮＡ表达随着吴茱萸碱浓度的增加和时间的延长其表达逐渐下降。当吴茱萸碱与ＤＤＰ联用时,可明显提高Ａ５４９／ＤＤＰ细胞对化疗药的敏感性,凋亡细胞显著增加（Ｐ＜０.０５）。Ｗｅｓｔｅｒｎｂｌｏｔ法结果提示Ａ５４９／ＤＤＰ细胞中ｐＩκＢ-α的表达水平随着吴茱萸碱作用时间的延长逐渐下降,ｐＩＫＫα表达则无显著变化。结论　吴茱萸碱可以通过抑制ＩκＢ-α的磷酸化阻断ＮＦ-κＢ信号通路,促进细胞凋亡,抑制细胞增殖,增加耐药细胞对ＤＤＰ的敏感性。     

LOX1在胃癌中的表达及其促肿瘤生长的机制探讨

目的:探讨血凝素样氧化型低密度脂蛋白受体1（lectin-like oxidized low density lipoprotein receptor-1,LOX1）在胃癌中的表达及与预后相关性,同时明确其促胃癌生长作用及潜在机制。方法:利用人类癌症基因组图谱(TCGA)、基因表达汇编(GEO)和癌症细胞系百科全书(CCLE)数据库分析LOX1在胃癌组织和细胞中的表达水平;利用多种统计方法分析LOX1表达水平与胃癌患者预后及临床特征的相关性;通过质粒转染敲低LOX1,进一步应用细胞功能实验（CCK-8、克隆形成、EdU实验）探究LOX1对胃癌细胞增殖能力的调控作用;通过Western blot、qPCR、免疫荧光分析LOX1对NF-κB信号通路激活的调控作用。结果:LOX1在胃癌组织和细胞中显著高表达;高水平LOX1提示胃癌患者预后较差且肿瘤较易发生转移;敲低LOX1显著抑制肿瘤细胞增殖;敲低LOX1显著抑制胃癌细胞中NF-κB信号通路的激活。结论:LOX1通过激活NF-κB信号通路对胃癌细胞增殖能力具有调控作用;LOX1可作为生物标记物提示患者预后;靶向LOX1可作为潜在胃癌治疗新方案。     

NF-κB inhibition in human hepatocellular carcinoma and its potential as adjunct to sorafenib based therapy

Nuclear factor-kappaB (NF-κB) has been shown to play an important role in the development and progression of cancer. In this study, we systematically examined NF-κBp65 signaling pathway in both human hepatocellular carcinoma (HCC) tissue and HCC cell lines. NF-κBp65 signaling pathway is aberrantly expressed and activated in both human HCC tissue and HCC Hep3B cells. Inhibition of NF-κB activity significantly reduced proliferation and invasion of Hep3B cells as well as down-regulated the expression of invasion-related molecules including matrix metalloproteinase (MMP)-2, MMP-9, membrane type-1 MMP (MT1-MMP), urokinase plasminogen activator (uPA) and vascular endothelial growth factor (VEGF). Hep3B cells exhibited a dose-dependent increase in apoptosis after receiving sorafenib treatment. Inhibition of NF-κB activity strongly sensitized Hep3B cells to sorafenib-induced cell death. Mechanistically, combined treatment of sorafenib and NF-κB inhibition enhanced inhibition of MAPK signaling and down-regulation of anti-apoptotic protein Mcl-1 expression. These observations indicate that inhibition of NF-κB may be a potential antineoplastic therapy for HCC, especially the combination of NF-κB inhibition and sorafenib provides a novel therapeutic strategy for patients with advanced-stage HCC.