白花蛇舌草上调miR-485-5p抑制人膀胱癌细胞系KU-19-19增殖、迁移和侵袭

目的 探讨白花蛇舌草对膀胱癌细胞系KU-19-19增殖、迁移和侵袭的影响和可能的分子机制.方法 将KU-19-19细胞分为对照组、白花蛇舌草组、miR-NC组、miR-485-5p组、anti-miR-NC组、anti-miR-485-5p组、白花蛇舌草+anti-miR-NC组、白花蛇舌草+anti-miR-485-...     

LncRNA DSCAM-AS1通过靶向miR-627-3p调控皮肤鳞状细胞癌增殖、侵袭和迁移

目的 探讨长链非编码RNA(LncRNA)唐氏综合征细胞粘附分子反义1(DSCAM-AS1)靶向miR-627-3p对人皮肤鳞状细胞癌增殖和侵袭能力的影响和分子机制.方法 采用实时荧光定量PCR(RT-qPCR)检测DSCAM-AS1和miR-627-3p在人永生化表皮细胞HaCaT和3种皮肤鳞状细胞癌细胞(SCC13...     

miR-660-5p靶向TET2促进乳腺癌的增殖、迁移和侵袭

目的 研究miR-660-5p/TET2通过PI3K/AKT/mTOR信号通路调节乳腺癌细胞增殖、凋亡、迁移和侵袭的机制。方法 收集2021年1月-2022年6月荆门市人民医院65例乳腺癌患者的乳腺癌组织和邻近正常组织样品,qPCR实验检测miR-660-5p在乳腺癌组织和细胞系中的表达水平,Kaplan-Meier生存分析和Log-rank检验分析miR-660-5p与患者临床病理特征的相关性;CCK8、流式细胞术和Transwell方法检测miR-660-5p对乳腺癌细胞增殖、凋亡、迁移和侵袭情况;双荧光素酶报告基因和蛋白质印迹实验明确miR-660-5p与TET2的关系;Western blot、CCK8和Transwell实验明确miR-660-5p通过靶向TET2调节乳腺癌的进程;Western blot实验检测miR-660-5p/TET2是否激活PI3K/AKT/mTOR信号通路;动物成瘤实验证明敲低miR-660-5p在乳腺癌中的作用。结果 miR-660-5p在乳腺癌组织中表达上调,且miR-660-5p的高表达与乳腺癌的淋巴结转移（P=0.003）,晚期TNM分期（P=0.009）和血管浸润（P=0.018）密切相关;敲低miR-660-5p可抑制乳腺癌细胞增殖、迁移和侵袭,诱导乳腺癌细胞凋亡;TET2是miR-660-5p的直接靶标,干扰TET2可以部分逆转敲低miR-660-5p对乳腺癌细胞恶性潜能的抑制作用;miR-660-5p下调TET2并激活PI3K/AKT/mTOR信号通路;敲低miR-660-5p可抑制体内肿瘤生长。结论 miR-660-5p通过靶向TET2和PI3K/AKT/mTOR信号通路来促进乳腺癌的进程,可能是治疗乳腺癌的潜在靶标。     

miR-101靶向调节c-Fos基因对卵巢癌SKOV3细胞增殖和侵袭的影响

目的探讨miR-101对卵巢癌SKOV3细胞增殖及侵袭能力的影响。方法将卵巢癌SKOV3细胞分为miR-101抑制剂组、miR-101模拟物组和对照组,分别将miR-101抑制剂或miR-101模拟物转染至miR-101抑制剂组或miR-101模拟物组细胞,对照组细胞转染无关序列,Real-time PCR法检测各组细胞miR-101表达水平,分别用MTT方法与Transwell细胞侵袭实验检测转染后各组SKOV3细胞的增殖与侵袭能力变化。Real-time PCR与Western blot实验分别检测各组细胞c-Fos mRNA和蛋白表达水平。结果转染miR-101模拟物后,SKOV3细胞miR-101的表达水平显著升高,SKOV3细胞的增殖与侵袭能力显著降低,c-Fos mRNA及蛋白的表达水平显著降低(P0.01);转染miR-101抑制剂后,SKOV3细胞miR-101的表达水平显著降低,SKOV3细胞的增殖与侵袭能力显著升高,c-Fos mRNA及蛋白的表达水平显著升高(P0.01)。结论 miR-101抑制卵巢癌SKOV3细胞的增殖和侵袭可能与下调c-Fos的表达相关。     

circ_POLA2靶向miR-30c-5p对胃癌细胞增殖、迁移及侵袭的影响

目的 ：探讨circ＿POLA2对胃癌细胞增殖、迁移及侵袭的影响及其可能作用机制。方法 ：qRT-PCR法检测胃癌组织与癌旁组织中circ＿POLA2、miR-30c-5p的表达量;Pearson法分析circ＿POLA2与miR-30c-5p在胃癌组织中表达量的相关性;体外培养人胃癌细胞HGC-27,根据转染物不同进行分组：si-NC组、si-circ＿POLA2组、miR-NC组、miR-30c-5p组、si-circ＿POLA2+NC-inhibitor组、si-circ＿POLA2+miR-30c-5p-inhibitor组;CCK-8法、Transwell实验依次检测细胞增殖及迁移、侵袭;双荧光素酶实验检测miR-30c-5p与circ＿POLA2的靶向关系;免疫印迹法检测蛋白PI3K、p-PI3K、Akt、p-Akt、MMP-2、MMP-9表达。结果 ：与癌旁组织比较,胃癌组织中circ＿POLA2的表达量升高,miR-30c-5p的表达量降低;circ＿POLA2与miR-30c-5p的表达呈负相关;与si-NC组比较,si-circ＿POLA2组miR-30c-5p的表...     

普鲁卡因调控CXCR7并影响AKT和STAT3信号通路从而抑制膀胱癌细胞活力、迁移和侵袭

目的:探讨普鲁卡因(PCA)及CXC趋化因子受体7(CXCR7)对膀胱癌细胞活力、迁移和侵袭的影响及潜在的作用机制。方法:用不同浓度PCA处理人膀胱癌RT4细胞,并将细胞分为PBS组(无PCA处理)、PCA组(4 mmol/L PCA处理)、siRNA阴性对照(si-Con)组(转染si-Con)、CXCR7 siRNA(si-CXCR7)组(转染si-CXCR7)、PCA+pcDNA组(4 mmol/L PCA处理并转染pcDNA)和PCA+pcDNA-CXCR7组(4 mmol/L PCA处理并转染pcDNACXCR7),siRNA和pcDNA均用脂质体法转染至RT4细胞。运用RT-qPCR检测RT4细胞中CXCR7的mRNA表达水平;采用CCK-8法检测各组细胞的活力;Transwell实验检测各组细胞的侵袭和迁移能力;Western blot检测各组细胞中CXCR7、AKT、STAT3、p-AKT和p-STAT3的蛋白水平。结果:相较于PBS组,不同浓度PCA处理后RT4细胞的活力、迁移能力和侵袭能力显著降低(P<0. 05),PCA组RT4细胞中CXCR7的mRNA和蛋...     

20(S)-人参皂苷Rg3抑制人急性单核细胞白血病THP-1细胞迁移和侵袭

目的：观察低极性20(S)-人参皂苷Rg₃(S-Rg₃)体外抑制人急性单核细胞白血病THP-1细胞迁移和侵袭的作用。方法：将不同浓度(0、20、40和80 mg/L)的S-Rg₃加入THP-1细胞培养体系,Transwell小室检测S-Rg₃抑制细胞迁移的作用。采用免疫细胞化学、免疫荧光、Western blot和RT-qPCR等多种方法,检测S-Rg₃对THP1细胞黏附、迁移和侵袭相关蛋白表达的影响及其相关机制。结果：S-Rg₃能有效地抑制THP-1细胞迁移,下调黏附相关蛋白N-cadherin,迁移相关蛋白C-X-C趋化因子受体4(CXCR4)和基质细胞衍生因子1(SDF-1),以及侵袭相关蛋白基质金属蛋白酶2(MMP2)的表达,但上调抑制迁移侵袭相关蛋白E-cadherin、金属蛋白酶组织抑制物1(TIMP1)和TIMP3表达,同时下调CXCR4 mRNA表达,上调E-cadherin和TIMP1 mRNA表达,呈剂量依赖性。此外,S-Rg_...     

IL-8通过激活PI3K/AKT通路促进结肠癌细胞增殖、迁移和侵袭

为研究IL-8对结肠癌细胞增殖、迁移和侵袭的影响并探讨其作用机制,应用免疫组织化学法检测结肠癌组织和正常癌旁组织中IL-8的表达;MTT法检测不同浓度IL-8处理或抑制PI3K/AKT信号通路后结肠癌细胞SW480的增殖情况;Western blotting检测IL-8处理后SW480细胞中p-AKT、t-AKT蛋白的表达;Transwell法检测抑制PI3K/AKT通路或IL-8处理后SW480细胞的迁移和侵袭能力。结果显示,与癌旁组织相比,结肠癌组织中IL-8的表达水平明显提高;IL-8处理后SW480细胞的增殖、迁移和侵袭能力明显增加,且PI3K/AKT通路异常激活;抑制PI3K/AKT通路可逆转IL-8对SW480细胞增殖、迁移和侵袭的促进作用。由此IL-8可促进结肠癌细胞增殖、迁移和侵袭,其机制可能与激活PI3K/AKT通路有关,可为IL-8在结肠癌中的治疗提供参考。     

下调miR-21 抑制PI3K / AKT 信号通路对白血病细胞增殖凋亡的影响

目的:探讨miR-21 对白血病K562 细胞增殖凋亡的影响及对PI3K/ AKT 信号通路的调控作用。方法:将K562 细胞分为对照组、miR-21 NC 组和miR-21 干扰组,对照组不做处理,后两组采用阳离子脂质体LipofectamineTM2000 转染miR-21 inhibitor 和miR-21 negative control。转染48 h 后,利用实时荧光定量(qRT-PCR)检测各组细胞中miR-21 mRNA 的表达情况,采用MTT 比色法检测miR-21 对细胞增殖率的影响,流式细胞仪检测miR-21 对细胞周期和凋亡率的影响,蛋白质免疫印迹法(Western blot)检测miR-21 对各组细胞中PI3K/ AKT 信号通路相关蛋白PI3K、AKT 和p-AKT 表达的影响。结果:miR-21 干扰组中细胞的miR-21 mRNA 表达水平和细胞的存活率较对照组和miR-21 NC 组均明显降低;与对照组和miR-21 NC 组比较,流式细胞仪检测miR-3 干扰组中G0/ G1 期所占细胞比例明显升高,S 期细胞所占比例明显下降,细胞的凋亡率明显升高。Western blot 检测p-AKT 的表达水平较对照组和miR-21 NC 组明显下降,但PI3K 和AKT 蛋白的表达水平变化不大。结论:下调miR-21 能够抑制白血病K562 细胞的增殖,促进其凋亡,其作用机制可以与抑制PI3K/ AKT 信号通路有关。     

敲除EZH2抑制人宫颈癌细胞系迁移和侵袭

目的 研究zeste基因同源蛋白2(EZH2)对人宫颈癌细胞系迁移和侵袭的影响,并初步探讨其作用机制.方法 利用CRISPR/Cas9敲除人宫颈癌HeLa和人子宫颈鳞癌SiHa细胞系中EZH2的表达.划痕实验检测细胞迁移;Transwell小室法检测细胞体外迁移和侵袭;实时荧光定量PCR检测STAT3 mRNA水平;W...     

CTHRC1 facilitates bladder cancer cell proliferation and invasion through regulating the PI3K/Akt signaling pathway

IntroductionEmerging evidence has illustrated that Collagen triple helix repeat containing 1 (CTHRC1) is crucial for tumorigenesis and development. However, the effects of CTHRC1 on bladder cancer progression remain largely unclear. Here, we aim to investigate the function and mechanism of CTHRC1 in behaviors of bladder cancer cells in vitro and in vivo.Material and methodsInterference assays were applied to determine the biological functions of CTHRC1. The expression of CTHRC1 was examined by quantitative real time-PCR (qRT-PCR), Western blot and immunohistochemical (IHC) analysis. Effects of CTHRC1 on proliferation, migration and invasion were evaluated by CCK-8, colony formation, flow cytometry, EdU staining, wound healing, transwell and western blot assays. Bladder cancer cells transfected with sh-CTHRC1 were injected into nude mice to explore the effect of CTHRC1 on tumorigenesis in vivo.ResultsCTHRC1 expression was increased in bladder cancer tissues and cell lines compared with normal controls, and associated with advanced clinical stage and lymph node metastasis. Also, patients with high levels of CTHRC1 expression were found to have a poor prognosis. Knockdown of CTHRC1 alleviated bladder cancer cell proliferation, migration and invasion in vitro and impeded tumorigenesis in vivo. Moreover, mechanistic investigation indicated that CTHRC1 could regulate the PI3K/Akt signaling pathway.ConclusionsOur data demonstrated that CTHRC1 played an oncogenic role in bladder cancer by modulating the PI3K/Akt signaling pathway, which sheds novel light on diagnosis and treatment of bladder cancer.     

miR-128-3p过表达靶向MAPK1抑制膀胱癌5637细胞株的侵袭，迁移和上皮间质转化

目的　研究miR-128-3p过表达对膀胱癌5637细胞株的侵袭,迁移和上皮间质转化影响。 方法　基因预测软件TargetScan筛选出miR-128-3p的靶基因,荧光素酶报告实验验证;RT-PCR检测miR-128-3p和MAPK1的表达,Transwell检测细胞侵袭情况,划痕实验检测细胞迁移能力,Western blot检测E-cadherin、N-cadherin、ERK1/2、c-Myc和c-fos的表达,免疫荧光检测Vimentin的表达;裸鼠皮下注射建立移植瘤模型,30 d后检测瘤重量,绘制存活曲线,检测移植瘤中Vimentin、miR-128-3p、MAPK1、ERK1/2、c-Myc和c-fos的量。 结果　miR-128-3p靶向抑制MAPK1表达;miR-128-3p过表达后,侵袭细胞数目、伤口愈合率降低,E-cadherin表达上调,N-cadherin表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。经miR-128-3p干预,裸鼠体内移植瘤重量减轻,存活率增加,miR-128-3p表达上调,MAPK1的表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。 结论　过表达miR-128-3p通过靶向抑制MAPK1表达来抑制膀胱癌细胞5637的侵袭能力、迁移能力、上皮-间充质转化和ERK1/2、c-Myc和c-fos通路。     

miR-128-3p过表达靶向MAPK1抑制膀胱癌5637细胞株的侵袭，迁移和上皮间质转化

目的　研究miR-128-3p过表达对膀胱癌5637细胞株的侵袭,迁移和上皮间质转化影响。 方法　基因预测软件TargetScan筛选出miR-128-3p的靶基因,荧光素酶报告实验验证;RT-PCR检测miR-128-3p和MAPK1的表达,Transwell检测细胞侵袭情况,划痕实验检测细胞迁移能力,Western blot检测E-cadherin、N-cadherin、ERK1/2、c-Myc和c-fos的表达,免疫荧光检测Vimentin的表达;裸鼠皮下注射建立移植瘤模型,30 d后检测瘤重量,绘制存活曲线,检测移植瘤中Vimentin、miR-128-3p、MAPK1、ERK1/2、c-Myc和c-fos的量。 结果　miR-128-3p靶向抑制MAPK1表达;miR-128-3p过表达后,侵袭细胞数目、伤口愈合率降低,E-cadherin表达上调,N-cadherin表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。经miR-128-3p干预,裸鼠体内移植瘤重量减轻,存活率增加,miR-128-3p表达上调,MAPK1的表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。 结论　过表达miR-128-3p通过靶向抑制MAPK1表达来抑制膀胱癌细胞5637的侵袭能力、迁移能力、上皮-间充质转化和ERK1/2、c-Myc和c-fos通路。     

胡椒碱对卵巢癌细胞株SKOV-3侵袭与凋亡的影响及其机制

目的:观察胡椒碱对卵巢癌细胞株SKOV-3的侵袭与凋亡的影响并讨论其作用机制.方法:CCK-8法检测不同浓度的胡椒碱(0,5,10和20μmol/L)对SKOV-3细胞作用不同时间(12,24和36 h)后,SKOV-3细胞存活率的变化;Transwell小室法与TUNEL法分别检测SKOV-3细胞侵袭能力与凋亡能力的变化;Western印迹检测p-PI3K与p-Akt蛋白表达水平的变化.结果:CCK-8法结果显示胡椒碱随浓度增加与培养时间增长,对SKOV-3细胞存活率的抑制程度增强;采用10μmol/L胡椒碱培养SKOV-3细胞36 h后,SKOV-3细胞的细胞侵袭数明显降低,细胞凋亡数明显增加,差异有统计学意义(P<0.05);p-PI3K与p-Akt蛋白表达水平明显降低,差异有统计学意义(P<0.05).结论:胡椒碱能够抑制卵巢癌细胞SKOV-3的增殖与侵袭能力,其增加凋亡能力可能与抑制PI3K/Akt信号通路有关.     

槲皮苷通过抑制PI3K/AKT信号通路诱导胃癌SGC7901细胞凋亡

目的:探讨槲皮苷是否通过抑制PI3K/AKT信号通路诱导人胃癌SGC7901细胞凋亡。方法:选取SGC7901细胞作为研究对象,采用MTT法检测槲皮苷对SGC7901细胞的毒性作用并测定IC50值。实验分为对照组(不加药处理)、槲皮苷组(采用200μmol/L槲皮苷处理)、PI3K/AKT通路激动剂胰岛素样生长因子1(IGF-1)组(采用100μg/L IGF-1处理)和槲皮苷+IGF-1组(采用200μmol/L槲皮苷+100μg/L IGF-1共处理)。处理48 h后,采用流式细胞术检测细胞凋亡,Western blot法检测cleaved caspase-3、p-AKT(Ser473)、AKT、p-PI3K(Tyr508)和PI3K的蛋白水平。结果:从100μmol/L开始,随着槲皮苷处理浓度的逐渐升高,SGC7901细胞活力显著降低(P 0. 05),槲皮苷作用48 h的IC50值为275. 40μmol/L。200μmol/L槲皮苷作用SGC7901细胞48 h后,与对照组比较,细胞凋亡率和cleaved caspase-3蛋白水平显著上升(P 0. 05),p-AKT和p-PI3K蛋白水平显著降低(P 0. 05),然而IGF-1与槲皮苷共同作用时,IGF-1可逆转槲皮苷对SGC7901细胞的作用效果。结论:槲皮苷能够诱导胃癌SGC7901细胞凋亡,其作用机制可能与抑制PI3K/AKT信号通路的激活有关。     

miR-451 inhibits invasion and proliferation of bladder cancer by regulating EMT

MicroRNAs (miRNAs) have recently been reported play a crucial role in some tumors. In order to investigate the association of miR-451 with bladder cancer, we investigate the expression of miR-451 in bladder cancer tissues and its role in biological behavior of T24, 5637 and J28 bladder cancer cell lines. Quantitative RT-PCR results showed miR-451 was significantly down-regulated in bladder cancer tissues and paracancerous tissues compared with normal bladder tissues. miR-451 expression was significantly associated with histological differentiation degree and TNM stage. Over-expression of miR-451 was established by transfecting miR-451 mimics into T24, 5637 and J28 cells, and its effects on the biological behavior of bladder cancer were studied using transwell assay, migration assay, adhesion assay, MTT and flow cytometry. Results indicated over-expression of miR-451 significantly inhibited cell proliferation, migration, invasion and induced apoptosis of the bladder cancer cells. Furthermore, we investigated the expression level of EMT related proteins in transfected 5637 cells by western blot. Results shown E-cadherin was up-regulated more significantly than N-cadherin, vimentin and Snail. N-cadherin and vimentin were up-regulated significantly when miR-451 was inhibited in miR-451 inhibitor group, however, no significant changes in mimics group. In conclusion, miR451 should be a tumor-suppressing gene in bladder cancer. miR-451 could maintain the bladder tumor cells in epithelial phenotype, inhibit EMT process, thereby reducing the invasion and migration of tumor cells.     

miR-204 inhibits invasion and epithelial-mesenchymal transition by targeting FOXM1 in esophageal cancer

MicroRNAs (miRNAs), endogenous noncoding small RNAs, have been reported to play crucial roles in epithelial-mesenchymal transition (EMT) in cancers. Deregulation of microRNA-204 (miR-204) has been documented in many cancers, but its role in the development of esophageal cancer (EC) has not been studied. Here, we reported the role of miR-204 in invasion and EMT in EC. We identified an inverse correlation between miR-204 expression level and the invasion and EMT phenotype of EC cells, and up-regulation of miR-204 inhibited invasion and EMT phenotype of EC cells. Furthermore, we showed that forkhead box protein M1 (FOXM1) was a direct target gene of miR-204, and miR-204 regulated invasion and EMT in EC by acting directly on the 3’UTR of FOXM1 mRNA and suppressing its protein expression. We also explored the anti-tumor effect of miR-204, and found that overexpression of miR-204 suppressed the growth of esophageal tumors in vivo. These findings suggest that miR-204 might be a suppressor of invasion and EMT in EC, which offers a novel potential therapeutic target for EC.     

Modulatory role of garlicin in migration and invasion of intrahepatic cholangiocarcinoma via PI3K/AKT pathway

Increasing evidences have indicated the role of garlicin in inhibiting the progression of various tumors including glioma, pulmonary carcinoma and pancreatic carcinoma, via mediating cell apoptosis or cell cycle. The regulatory effect and related molecular mechanism of garlicin in intrahepatic cholangiocarcinoma, however, remained unknown. This study thus aimed to investigate this scientific issue. HCCC-9810 cell line was treated with serially diluted garlicin, followed by cell proliferation assay using MTT approach. Transwell migration and invasion assays were further employed the regulatory effect of garlicin. The expression level of p-AKT and AKT proteins in tumor cells was quantified by Western blot. The growth of tumor cells was significantly inhibited by high concentration of garlicin (> 1.5 μM). Lower concentration of garlicin showed dose-dependent inhibition of tumor cell invasion and migration. After using specific agonist IGF-1 (50 ng/mL) of PI3K/AKT signaling pathway, such facilitating effects of garlicin were depressed (P < 0.05). Western blotting showed significantly decreased phosphorylation level of AKT after treated with gradient concentrations of garlicin, while leaving the total AKT protein level unchanged. Garlicin may inhibit the invasion and migration of intrahepatic cholangiocarcinoma cells via inhibiting PI3K/AKT signaling pathway.     

miR-551b-3p在胃癌组织中低表达并抑制胃癌细胞系HGC-27的增殖、迁移和侵袭

目的探讨miR-551b-3p在胃癌中的表达变化情况,及其对胃癌细胞功能的影响。方法用real-time PCR的方法检测60例胃癌组织及其对应的癌旁组织中miR-551b-3p的表达量,使用miR-551b-3p模拟物(miR-551b-3p mimic)转染胃癌细胞系HGC-27,CCK-8法检测细胞增殖;划痕愈合实验检测细胞迁移;Transwell法检测细胞侵袭。结果 1)miR-551b-3p在胃癌癌症组织中的表达明显低于癌旁组织(P0.05)。2)过表达miR-551b-3p mimic可以明显减弱胃癌细胞系HGC-27的增殖、迁移和侵袭能力。结论 miR-551b-3p在胃癌组织中低表达,并且抑制胃癌细胞系HGC-27的增殖、迁移和侵袭,可能与胃癌发生发展密切相关。