嗜酸乳杆菌、长双歧杆菌上清液对HCT116、SW480细胞增殖的影响

目的:研究嗜酸乳杆菌(ATCC 4356)与长双歧杆菌(ATCC 15707)上清液对HCT116、SW480人结肠癌细胞增殖作用的影响。方法:体外培养HCT116、SW480、嗜酸乳杆菌和长双歧杆菌,分别提取嗜酸乳杆菌和长双歧杆菌的上清液。通过MTT法分析嗜酸乳杆菌和长双歧杆菌不同组分对人结肠癌细胞增殖的影响。结果:嗜酸乳杆菌上清液对SW480细胞抑制作用弱于MRS培养基(F=7.24,P=0.043 3);长双歧杆菌上清液与BHI培养基对SW480细胞抑制作用的差异无统计学意义(F=1.81,P=0.236 6),而不同浓度之间细胞抑制率的差异有统计学意义(F=18.07,P=0.003 2);嗜酸乳杆菌上清液对HCT116细胞抑制作用强于MRS培养基(F=10.33,P=0.023 6),而不同浓度之间细胞抑制率的差异无统计学意义(F=3.51,P=0.097 3); BHI培养基对HCT116细胞促增殖作用强于长双歧杆菌上清液(F=41.12,P=0.001 4)。结论:嗜酸乳杆菌上清液在一定的浓度范围内对HCT116细胞增殖具有抑制作用,对SW480细胞无抑制作用。长双歧杆菌...     

野黄芩苷调控Wnt/β-catenin信号通路对结肠癌细胞迁移和侵袭的影响

目的 观察野黄芩苷(Scu)对人结肠癌细胞迁移和侵袭的影响及相关作用机制。方法 采用CCK8法检测40、80、120、160、200、240、280μmol·L^-1 Scu对人结肠癌SW480和HCT116细胞以及人肾皮质近曲小管上皮HK-2细胞活力的影响，计算半数抑制浓度(IC50)值。采用细胞划痕实验、Transwell细胞迁移和小室侵袭实验检测Scu对SW480和HCT116细胞迁移和侵袭的影响，Western blot法检测50、100、150μmol·L^-1 Scu对SW480细胞迁移、侵袭、上皮-间质转化(EMT)和Wnt/β-catenin信号通路相关蛋白表达的影响。结果 120、160、200、240、280μmol·L^-1Scu显著抑制SW480和HCT116细胞的活力(P <0.01)，作用24 h后的IC50值分别为157.7和179.2μmol·L^-1。与空白组比较，50、100、150μmol·L^-1 Scu组SW480和HCT116细胞划痕宽度...     

苦参碱和氧化苦参碱抗大肠癌药理作用研究进展

苦参碱和氧化苦参碱能防治氧化偶氮甲烷/葡聚糖硫酸钠诱发小鼠原发性结直肠癌和CT26细胞或SW480-EGFP细胞移植瘤在大鼠或小鼠体内生长。体外实验发现苦参碱和氧化苦参碱能浓度相关地抑制大肠癌SW480细胞、SW480-EGFP细胞、SW480/M5细胞、SW620细胞、SW1116细胞、LoVo细胞、HT-29细胞、Colon26细胞、HCT116细胞、HCT-8细胞、HCL细胞和LS174t细胞增殖,并诱导凋亡。苦参碱能增强奥沙利铂抗SW480细胞、SW620细胞和SW1116细胞的作用。氧化苦参碱能增强5-氟尿嘧啶抗LoVo细胞的作用。     

小檗碱抗肿瘤作用与Wnt/-βcaten in信号转导关系

目的证明小檗碱抗肿瘤作用机制可能与信号转导过程的调控有关。方法采用细胞增殖抑制和Hoechst 33258染色凋亡实验比较小檗碱和黄连总碱对人结肠癌HCT116和SW 480细胞的作用。利用Tcf-4报告基因研究小檗碱对肿瘤细胞的信号转导影响。结果小檗碱在5~40 mg.L-1浓度范围内呈浓度依赖性和时间依赖性抑制人结肠癌HCT116和SW 480细胞的增殖;小檗碱(20 mg.L-1)处理72 h后的HCT116和SW 480细胞出现明显凋亡;相当于小檗碱浓度的黄连总碱有类似于小檗碱的作用。20~40 mg.L-1小檗碱和黄连总碱均能明显抑制-βcaten in/Tcf介导的转录活性。结论黄连总碱的抗肿瘤作用可能与其主要成分小檗碱有关;其抗肿瘤作用机制至少与抑制W nt/-βcaten in信号通路有关。     

lncRNA FOXCUT调控Notch通路抑制结肠癌细胞增殖、侵袭的实验研究#br#

目的　研究长链非编码RNA（lncRNA）FOXC1启动子临近转录体（FOXCUT）对结肠癌细胞（HCT116）增殖、侵袭的影响及作用机制。方法　向HCT116细胞转染FOXCUT siRNA（FOXCUT siRNA组）,以转染阴性siRNA为阴性对照组（FOXCUT siRNA-NC组）,未转染组为正常对照组（NC组）,通过qPCR法确定FOXCUT干扰效果,并检测结肠癌及癌旁组织、人结肠癌细胞株（SW480、SW620、HCT116、HT29）和人正常结肠上皮细胞（NCM460）中FOXCUT mRNA表达水平;通过CCK-8、集落形成实验和Transwell实验检测沉默FOXCUT对HCT116细胞增殖、集落形成能力及侵袭能力的影响;qPCR检测沉默FOXCUT后HCT116细胞中Notch1、Hes1 mRNA表达变化;蛋白免疫印迹（Western blot）实验检测HCT116细胞中Notch1、Hes1蛋白表达情况。结果　与癌旁组织比较,结肠癌组织中FOXCUT mRNA表达水平显著增高（P＜0.01）;与NCM460细胞比较,SW480、SW620、HCT116、HT29结肠癌细胞株中FOXCUT mRNA表达水平均显著增高（P＜0.05）,其中以HCT116细胞中表达水平最高;与FOXCUT siRNA-NC组比较,FOXCUT siRNA组HCT116细胞中FOXCUT mRNA表达水平显著降低（P＜0.01）;NC组、FOXCUT siRNA-NC组HCT116细胞中FOXCUT mRNA表达水平差异无统计学意义（P＞0.05）;沉默FOXCUT可显著抑制细胞增殖、集落形成能力及侵袭（P＜0.05）,并抑制Notch1、Hes1 mRNA及蛋白表达（P＜0.01）。结论　沉默FOXCUT可抑制结肠癌HCT116细胞增殖及侵袭,可能与抑制Notch通路有关。     

LncRNA TMPO-AS1调节miR-340-5p/RUNX1轴对结直肠癌细胞增殖、迁移和侵袭的影响

目的 探讨长链非编码RNA TMPO反义RNA1(LncRNA TMPO-AS1)调控miR-340-5p/RUNT相关转录因子1(RUNX1)轴对结直肠癌(CRC)细胞增殖、迁移和侵袭的影响。方法 qRT-PCR检测人正常结肠上皮细胞HCoEpiC、人CRC细胞SW480、HCT116、LOVO、HT-29中TMPO-AS1、miR-340-5p、RUNX1表达;将HCT116细胞随机分为：si-ctrl组、si-TMPO-AS1组、mimics NC组、miR-340-5p组、si-TMPO-AS1+anti-miR-ctrl组、si-TMPO-AS1+anti-miR-340-5p组;qRT-PCR检测6组HCT116细胞TMPO-AS1、miR-340-5p、RUNX1水平;CCK-8法检测HCT116细胞活力;EDU法检测HCT116细胞增殖;Transwell检测HCT116细胞迁移和侵袭;western blot检测HCT116细胞RUNX1、PCNA、MMP-2表达;双荧光素酶报告基因实验分别验证TMPO-AS1和miR-340-5p、miR-340-5p和RUNX1的...     

西红花苷抑制NFATc3活性促进与肿瘤成纤维细胞共培养的结直肠癌细胞放射敏感性

目的 探究西红花苷对与肿瘤成纤维细胞（CAFs）共培养的结直肠癌细胞放射敏感性的影响,并探究分子机制。方法 通过实时荧光定量聚合酶链式反应（qRT-PCR）和蛋白质免疫印记（Western blotting）比较人正常结肠上皮细胞NCM460和结直肠癌细胞系SW480、SW620、HCT116、LOVO中活化T细胞核因子c3（NFATc3）表达差异。建立CAFs与HCT116细胞共培养体系,西红花苷处理细胞后经不同剂量（0、2、4、6、8 Gy）X射线照射,细胞克隆形成实验分析细胞存活分数（SF）。再将HCT116细胞分为对照组、6 Gy组、CAFs/HCT116＋6 Gy组、CAFs/HCT116＋西红花苷＋6 Gy组,进行对应处理后,MTT法检测各处理组HCT116细胞活性,流式细胞术检测各处理组HCT116细胞凋亡率,Hoechst 33258染色观察各处理组HCT116细胞凋亡形态,qRT-PCR和Western blotting测定各处理组HCT116细胞中NFATc3表达水平。结果 相较于人正常结肠上皮细胞NCM460,人结直肠癌细胞系SW480、SW620、HCT116、LOVO中的NFATc3 mRNA和蛋白相对表达量均显著上调（P＜0.05）。与HCT116细胞比较,与CAFs共培养的HCT116细胞经辐射后的SF显著增高（P＜0.05）;与CAFs共培养的HCT116细胞比较,与CAFs共培养的HCT116细胞经西红花苷处理再进行辐射的SF显著降低（P＜0.05）。与6 Gy组比较,CAFs/HCT116＋6 Gy组HCT116细胞存活率显著升高（P＜0.05）,细胞凋亡率显著下降（P＜0.05）,细胞内染色也较为均匀,未见致密浓染的凋亡状细胞,细胞中NFATc3 mRNA和蛋白相对表达量均显著上调（P＜0.05）;而与CAFs/HCT116＋6 Gy组比较,CAFs/HCT116＋西红花苷＋6 Gy组HCT116细胞存活率显著降低（P＜0.05）,细胞凋亡率显著升高（P＜0.05）,细胞中有较多致密浓染、碎裂荧光块,同时,细胞中NFATc3 mRNA和蛋白相对表达量显著下调（P＜0.05）。结论 西红花苷能够明显提高与CAFs共培养的结直肠癌HCT116细胞的放射敏感性,从而促进细胞发生凋亡,该作用可能与抑制NFATc3有关。     

左金丸联合西妥昔单抗诱导KRAS突变肠癌细胞铁死亡作用研究

目的:探讨左金丸(ZJW)对KRAS突变的人结肠癌细胞西妥昔单抗(CET)抵抗的效果及机制。方法:采用Sanger测序法检测SW620、Lovo、HCT116、HT29和Caco2细胞KRAS基因突变状态；采用CCK-8法检测ZJW、CET、ZJW联合CET以及联合不同细胞死亡抑制剂对上述细胞存活率的影响，并观察ZJW联合CET对KRAS突变型细胞(SW620、Lovo及HCT116)耐药性的影响；流式细胞术、比色法和FerroOrange荧光探针检测和观察ZJW、CET以及它们联合作用对KRAS突变型细胞内ROS、GSH及Fe²⁺水平的影响；Western blot和Real-time PCR法检测ZJW联合CET对铁死亡相关通路蛋白和mRNA的调控作用。结果:KRAS突变型细胞为HCT116(KRAS^G13D),Lovo(KRAS^G13D),SW620(KRAS^G12V),KRAS野生型细胞为HT29和Caco2;与125μg/mL CET处理组比较，相同浓度CET明显降低KRAS野生型细胞Ca...     

微小RNA-152-3p 对结肠癌细胞增殖、迁移和侵袭的影响及其作用机制研究

目的探讨微小RNA(miR)-152-3p对结肠癌细胞的增殖、迁移和侵袭的影响及机制。方法2019年2月至2020年4月,将结肠癌SW480细胞分为miR-152-3p模拟物阴性对照(miR-NC)组、miR-152-3p模拟物(miR-152-3p)组、抑制物(anti-miRNC)组、miR-152-3p抑制物(anti-miR-152-3p)组、阴性对照(si-NC)组、沉默转移相关蛋白2(si-MTA2)组、miR-152-3p模拟物+空载体(miR-152-3p+pcDNA3.1)组、miR-152-3p模拟物+过表达MTA2(miR-152-3p+pcDNA3.1-MTA2)组,均用脂质体法转染至结直肠癌SW480细胞中,另取未转染结直肠癌SW480细胞记为Control组。采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测miR-152-3p和MTA2 mRNA的表达水平;蛋白质印迹法测定蛋白的表达;MTT法检测细胞活性;Transwell检测细胞迁移和侵袭;双荧光素酶报告基因检测实验检测荧光活性。结果结肠癌HCT116、SW480及LoVo细胞中miR-152-3p表达分别为0.72±0.04、0.29±0.02、0.49±0.01,人正常结肠上皮细胞NCM460中miR-152-3p表达为1.06±0.12,与正常结肠上皮细胞NCM460相比,结肠癌细胞HCT116、LoVo、SW480中MTA2 mRNA和蛋白较高,miR-152-3p较低(P<0.05);Control组、miR-NC组、miR-152-3p组、si-NC组及si-MTA2组结肠癌SW480细胞吸光度分别为0.92±0.22、0.96±0.17、0.31±0.07、0.99±0.17及0.32±0.09,细胞迁移数分别为(172.00±23.52)个、(169.00±20.66)个、(53.67±12.22)个、(155.67±16.8)个及(54.33±8.74)个,细胞侵袭数分别为(124.67±10.02)个、(122.33±9.45)个、(26.00±5.00)个、(108.33±10.02)个及(42.00±4.00)个,与miR-NC 组相比,miR-152-3p 组SW480细胞中细胞周期蛋白D1(cyclin D1)、基质金属蛋白酶(MMP)-2及SW480细胞活性、迁移和侵袭数量均较低,周期素依赖激酶抑制剂p21(P21)及上皮钙黏素(E-cadherin)较高(P<0.05);与si-NC组相比,si-MTA2组SW480细胞中cyclin D1、MMP-2及SW480细胞活性、迁移和侵袭数量均较低,P21及E-cadherin较高(P<0.05)。转染miR-152-3p和MTA2野生型表达载体的结肠癌SW480细胞荧光素酶活性显著降低(P<0.05)。相比miR-152-3p+pcDNA3.1组,miR-152-3p+pcDNA3.1-MTA2组SW480细胞中P21、E-cadherin的表达较低,MTA2、cyclin D1、MMP-2蛋白的表达较高,SW480细胞活性、迁移、侵袭数量较高(P<0.05)。结论miR-152-3p可能通过下调MTA2抑制结肠癌细胞的增殖、迁移和侵袭。     

结直肠癌中Kiss-1基因启动子甲基化与Kiss-1基因表达的关系

目的：研究结直肠癌中Kiss-1基因启动子甲基化状态对Kiss-1基因表达的影响。方法：应用甲基化特异性PCR （MSP）方法检测73例结直肠癌、正常结直肠组织和人结直肠癌细胞HCT116、SW480、W1116、LoVo中Kiss-1基因启动子甲基化状态,应用realtime-PCR、Western-blot技术检测相应组织和细胞中Kiss-1基因mRNA和蛋白质（Metastine）的表达量。结果：结直肠癌中Kiss-1基因甲基化阳性率（82.19%）高于正常组织（6.31%）（PSW480〉SW1116〉HCT116,差异有统计学意义（p〈0.05）。结论：结直肠癌中Kiss-1基因启动子甲基化可能引起Kiss-1基因表达下调。     

长链非编码RNA赖氨酰氧化酶样蛋白1反义RNA在结直肠癌中的表达及对结直肠癌细胞增殖、侵袭和迁移能力的影响

目的 探究长链非编码RNA(lncRNA)赖氨酰氧化酶样蛋白1反义RNA(LOXL1-AS1)在结直肠癌中的表达及对结直肠癌细胞增殖、侵袭及迁移能力的影响和可能的作用机制。方法 选取2016年1月至2018年12月在河南省人民医院行结直肠癌根治术的56例病人新鲜癌组织,采用实时定量聚合酶链反应(qRT-PCR)检测结直肠癌组织样本及结直肠癌细胞系SW480、SW620、HCT116和Lovo细胞中lncRNA LOXL1-AS1的表达水平,以配对癌旁组织及人正常结直肠黏膜细胞FHC作正常对照;构建靶向LOXL1-AS1序列的小干扰RNA(si-LOXL1-AS1)并转染SW480和Lovo细胞,采用细胞计数试剂盒(CCK-8)、平板克隆实验、划痕实验和Transwell小室法检测沉默lncRNA LOXL1-AS1表达对结直肠癌细胞增殖、侵袭和迁移能力的影响。采用蛋白质印迹法(Western blotting)检测沉默LOXL1-AS1表达后磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路相关蛋白及上皮-间质转化(EMT)相关蛋白的表达情况。结果 LOXL1-AS1在结直肠癌...     

SPNS2 promotes the malignancy of colorectal cancer cells via regulating Akt and ERK pathway

Colorectal cancer (CRC) is a prevalent malignant tumour that causes considerable cancer‐related deaths globally. The sphingolipid transporter 2 (SPNS2), a sphingosine‐1‐phosphate (S1P) transporter, modulates multiple biological events including malignancy of cancer cells. In this study, the effects of SPNS2 on CRC progression were studied. We found that SPNS2 expression was significantly upregulated in CRC tissues compared to that in adjacent non‐tumour tissues. To assess the role of SPNS2 in CRC cells, we performed loss‐ and gain‐of‐function experiments in SW480 and HCT116 cells, respectively. The results demonstrated that SPNS2 promoted proliferation, migration and invasion, and inhibited apoptosis in CRC cells. Additionally, SPNS2 enhanced the release of intracellular S1P, and increased S1P receptor 1 (S1PR1) and S1PR3 expression. Moreover, SPNS2 activated the Akt and ERK pathways, and the biological behaviours of SPNS2 were attenuated by Akt or ERK inhibitor in HCT116 cells. In conclusion, our results demonstrated that SPNS2 promoted proliferation, migration and invasion, and inhibited apoptosis by regulating S1P/S1PR1/3 axis and activating Akt and ERK pathway in CRC cells.     

m6A识别蛋白HuR调控lncRNA TRG-AS1抑制结直肠癌生长的机制研究

目的 探讨N6-甲基腺苷（m6A）识别蛋白人类抗原R(HuR)调控长链非编码RNA T细胞受体γ位点反义RNA 1(lncRNA TRG-AS1)对结直肠癌（CRC）生长的影响。方法 比色法检测CRC患者的癌组织、癌旁组织及正常结肠上皮细胞NCM460及CRC细胞HCT116、SW480、LOVO中m6A含量;实时荧光定量PCR(qPCR)检测TRG-AS1表达;Western blot检测HuR蛋白表达。将HCT116细胞分为Ct组、OE-NC组、OE-HuR组、si-NC组、si-HuR组、siHuR+pcDNA组、si-HuR+pcDNA-TRG-AS1组,CCK-8法检测细胞增殖;平板克隆实验检测细胞克隆形成能力;流式细胞术检测细胞凋亡率;划痕愈合实验检测细胞迁移;Transwell检测细胞侵袭;裸鼠体内移植瘤实验观察肿瘤生长情况;采用甲基化RNA免疫共沉淀（MeRIP）检测TRG-AS1上是否存在m6A位点;RNA pull-down实验和RNA免疫共沉淀（RIP）检测TRG-AS1与HuR蛋白的相互作用。结果 在CRC组织和细胞中,HuR蛋白、TRG-AS1高表达,m6A含...     

Rotenone restrains the proliferation,motility and epithelial–mesenchymal transition of colon cancer cells and the tumourigenesis in nude mice via PI3K/AKT pathway

Rotenone, a toxic rotenoid compound, has anti-tumour effects on several cancers. This study aims to clarify the effect of rotenone on the proliferation, apoptosis, invasion and migration of colon cancer cells and tumourigenesis in nude mice. The present results show that rotenone significantly inhibited the proliferation, promoted the apoptosis, and suppressed the invasion and migration of colon cancer cells in a dose-dependent manner. Rotenone inhibited PI3K/AKT pathway in LoVo and SW480 cells in a dose-dependent manner. In addition, rotenone regulated the proliferation, apoptosis, invasion, migration and EMT of LoVo and SW480 cells through PI3K/AKT pathway. In colon cancer xenograft mice, rotenone inhibited tumour volume and weight in nude mice, inhibited PI3K/AKT pathway and EMT in vivo. Therefore, rotenone inhibited the proliferation, invasion and migration, promoted the apoptosis of colon cancer cells through PI3K/AKT pathway in vitro, and suppressed the tumourigenesis in nude mice in vivo.     

Inhibition of microRNA‐21‐3p suppresses proliferation as well as invasion and induces apoptosis by targeting RNA‐binding protein with multiple splicing through Smad4/extra cellular signal‐regulated protein kinase signalling pathway in human colorectal cancer HCT116 cells

MicroRNA‐21‐3p (miR‐21‐3p), the passenger strand of pre‐mir‐21, has been found to be high‐expressing in various cancers and to be associated with tumour malignancy, which is proposed as a novel focus in malignant tumours. Colorectal cancer (CRC), currently known as one of the most prevalent malignancy, is a leading cause of cancer death. This study aimed to investigate the key role of miR‐21‐3p in CRC by inhibiting its expression using transfection with miR‐21‐3p inhibitors into human CRC HCT116 cells. Results showed that the expression of miR‐21‐3p was higher than other CRC cells used in the study including Lovo, HT29, Colo320 and SW480 cells, inhibition of which suppressed the proliferation and induced cell cycle arrest in HCT116 cells. Besides, transfection with miR‐21‐3p inhibitors also attenuated cell migration and invasion, and induced apoptosis as well. Moreover, luciferase assay confirmed RBPMS as a direct target of miR‐21‐3p in HCT116 cells. Further, miR‐21‐3p inhibitors increased the nuclear accumulation of Smad4 and reduced phosphorylation of ERK. Interestingly, we found that silence of RBPMS using RNA interference (siRNA) not only elevated the cell viability but also increased the phosphorylation of ERK and reversed the nuclear accumulation of Smad4 induced by miR‐21‐3p inhibitors in HCT116 cells. Data suggest that inhibition of miR‐21‐3p suppresses cell proliferation, invasion as well as migration and induces apoptosis by directly targeting RBPMS through Smad4/ERK signalling pathway in HCT116 cells. Our study demonstrates miR‐21‐3p as a potent target for suppressing tumour progression of CRC which may have implications in CRC therapy in the future.     

New benzimidazole acridine derivative induces human colon cancer cell apoptosis in vitro via the ROS-JNK signaling pathway

Aim:

To investigate the mechanisms underlying anticancer action of the benzimidazole acridine derivative N-{(1H-benzo[d]imidazol-2-yl)methyl}-2-butylacridin-9-amine(8m) against human colon cancer cells in vitro.

Methods:

Human colon cancer cell lines SW480 and HCT116 were incubated in the presence of 8m, and then the cell proliferation and apoptosis were measured. The expression of apoptotic/signaling genes and proteins was detected using RT-PCR and Western blotting. ROS generation and mitochondrial membrane depolarization were visualized with fluorescence microscopy.

Results:

8m dose-dependently suppressed the proliferation of SW480 and HCT116 cells with IC50 values of 6.77 and 3.33 μmol/L, respectively. 8m induced apoptosis of HCT116 cells, accompanied by down-regulation of Bcl-2, up-regulation of death receptor-5 (DR5), truncation of Bid, cleavage of PARP, and activation of caspases (including caspase-8 and caspase-9 as well as the downstream caspases-3 and caspase-7). Moreover, 8m selectively activated JNK and p38 without affecting ERK in HCT116 cells. Knockout of JNK1, but not p38, attenuated 8m-induced apoptosis. In addition, 8m induced ROS production and mitochondrial membrane depolarization in HCT116 cells. Pretreatment with the antioxidants N-acetyl cysteine or glutathione attenuated 8m-induced apoptosis and JNK activation in HCT116 cells.

Conclusion:

The new benzimidazole acridine derivative, 8m exerts anticancer activity against human colon cancer cells in vitro by inducing both intrinsic and extrinsic apoptosis pathways via the ROS-JNK1 pathway.     

红景天苷通过抑制STAT3活化调控结肠癌细胞增殖和转移

目的：研究红景天苷对结肠癌细胞增殖和转移的抑制作用,并探讨其分子机制。方法：采用平板克隆和CCK-8法检测结肠癌HCT116细胞活力,Transwell观察细胞侵袭与迁移能力,蛋白质印迹法检测增殖蛋白[c-Myc,细胞周期蛋白D1（cyclinD1）]、侵袭转移相关蛋白[E-钙黏附蛋白（E-cadherin）和基质蛋白酶9（MMP-9）]及JAK2/STAT3信号通路蛋白[STAT3、磷酸化STAT3（p-STAT3）、JAK2和磷酸化JAK2（p-JAK2）]的表达。结果：平板克隆和CCK-8实验显示,与对照组相比,0.5,1,2 μg·mL^-1红景天苷显著抑制HCT116细胞的增殖,且在该范围内有浓度依赖性。Transwell实验显示,0.5,1,2 μg·mL^-1红景天苷分别作用于HCT116细胞24 h,细胞的侵袭、转移能力也逐渐降低（P<0.05）。蛋白质印迹法检测结果显示：与对照组相比,0.5,1,2 μg·mL^-1红景天苷组c-Myc、cyclinD1、MMP-9、p-STAT3、p-JAK2蛋白水平均明显降低（P<0.05）,E-cadherin蛋白水平均升高（P<0.05）,而JAK2、STAT3蛋白水平无明显差异（P>0.05）。与红景天苷组比较,红景天苷+Y705D组细胞增殖能力增强,迁移和侵袭细胞数增多（P<0.05）,p-STAT3,c-Myc,cyclinD1,MMP-9蛋白表达增加,E-cadherin蛋白表达减少（P<0.05）;而与Y705D组比较,红景天苷+Y705D组细胞增殖能力减弱（P<0.05）,迁移和侵袭细胞数减少（P<0.05）,p-STAT3、c-Myc、cyclinD1、MMP-9蛋白表达减少（P<0.05）,E-cadherin蛋白表达增加（P<0.05）。结论：红景天苷抑制结肠癌细胞增殖和转移,其作用机制与抑制STAT3信号通路有关。     

Different involvement of DNA methylation and histone deacetylation in the expression of solute-carrier transporters in 4 colon cancer cell lines

The purpose of this study on the involvement of epigenetic control of the expression of solute carrier (SLC) transporters by DNA methylation and histone deacetylation in 4 colon cancer cells is to find the epigenetic control mechanisms of drug transporters in colon cancers. Human colon cancer cell lines (HCT116, HT29, SW48, SW480) were treated with 5-aza-2'-deoxycytidine (DAC), as a DNA methyltransferase inhibitor, followed by trichostatin A (TSA), as a histone deacetylase inhibitor. The mRNA expression and DNA methylation of several SLC transporters were analyzed by real-time polymerase chain reaction (PCR) and methylation-specific PCR, respectively. Among 12 SLC transporters possessing cytosine-phosphate-guanine (CpG) islands, thiamine transporter 2 (THTR2) (SLC19A3) gene showed a correlation between its mRNA expression level and DNA methylation status. TSA treatment increased histone H3 acetylation of THTR2 promoter region in all 4 colon cancer cell lines examined. HCT116 and SW48 cells showed a lack of THTR2 mRNA expression and methylation of its promoter, and DAC treatment induced its re-expression. In addition, the co-treatment with DAC and TSA increased THTR2 mRNA expression more markedly than DAC treatment in HCT116 and SW48 cells. In HT29 and SW480 cells that showed little methylation of THTR2 promoter, TSA treatment induced THTR2 mRNA expression markedly, but DAC treatment did not. In the 4 colon cancer cells examined, THTR2 mRNA expression is down-regulated by DNA methylation and/or histone deacetylation.