1文献来源 Becker A, Crombag L, Heideman DA, et al. Retreatment with Erlotinib: Regain of TKI sensitivity following a drug holiday for patients with NSCLC who initially responded to EGFR-TKI treatment [J]. Eur J Cance...1文献来源 Becker A, Crombag L, Heideman DA, et al. Retreatment with Erlotinib: Regain of TKI sensitivity following a drug holiday for patients with NSCLC who initially responded to EGFR-TKI treatment [J]. Eur J Cancer, 2011,47(17) :2603-2606.2展开更多
目的:观察微小核糖核酸-134-5p(mi R-134-5p)转染对宫颈癌细胞增殖和凋亡的影响,验证其可能的分子机制。方法:收集湖北医药学院附属人民医院肿瘤中心2016年5月至8月收治的8名宫颈癌患者肿瘤组织和相应癌旁组织。利用lipofectamine 2000...目的:观察微小核糖核酸-134-5p(mi R-134-5p)转染对宫颈癌细胞增殖和凋亡的影响,验证其可能的分子机制。方法:收集湖北医药学院附属人民医院肿瘤中心2016年5月至8月收治的8名宫颈癌患者肿瘤组织和相应癌旁组织。利用lipofectamine 2000将mi R-134-5p mimics转染至宫颈癌Hela和Si Ha细胞。采用MTT法和集落形成实验检测细胞增殖活性;流式细胞术(FCM)检测细胞周期和细胞凋亡;q RT-PCR检测宫颈癌组织和细胞mi R-134-5p m RNA表达以及宫颈癌细胞EGFR m RNA表达;Western blotting检测宫颈癌细胞EGFR信号通路相关蛋白的表达。结果:宫颈癌组织mi R-134-5p m RNA表达显著低于癌旁组织(P<0.01)。和转染mi R-NC的Hela和Si Ha细胞比较,转染mi R-134-5pmimics的宫颈癌Hela和Si Ha细胞mi R-134-5p m RNA表达显著升高;细胞增殖能力显著降低(转染第5天,Hela细胞:1.06±0.13 vs 1.32±0.07;Si Ha细胞:1.12±0.10 vs 1.42±0.12,均P<0.05);形成的集落数减少;G0/G1期细胞比例显著上升,S期和G2/M期细胞比例显著下降;细胞凋亡率显著增加[Hela细胞:(26.53±13.48)%vs(3.25±1.74)%;Si Ha细胞:(30.49±12.04)%vs(5.10±2.86)%,均P<0.05];EGFR m RNA和EGFR蛋白表达显著下调,其中EGFR m RNA,Hela细胞下调58%(P<0.01),Si Ha细胞下调41%(P<0.05);EGFR下游靶蛋白p-AKT、p-ERK1/2和Cyclin D1蛋白及p EGFR蛋白表达显著下调。结论:mi R-134-5p可显著抑制宫颈癌细胞增殖并促进细胞凋亡,其可能的分子机制是通过抑制EGFR基因的表达,抑制EGFR通路的活化。展开更多
Skin toxicity is a common symptom of anti-epidermal rowth factor receptor(EGFR) antibody treatment and is also a predictive marker of its efficacy in colorectal cancer patients. However, severe skin disorders induced ...Skin toxicity is a common symptom of anti-epidermal rowth factor receptor(EGFR) antibody treatment and is also a predictive marker of its efficacy in colorectal cancer patients. However, severe skin disorders induced by such antibodies negatively impact on the quality of life of patients and decreases drug compliance during treatment. If we can predict the high-risk group susceptible to severe skin toxicity before treatment, we can undertake the early management of any arising skin disorders and formulate a more accurate prognosis for anti-EGFR antibody treatment. Previous studies have identified molecular markers of skin toxicity induced by anti-EGFR antibody, such as EGFR polymorphisms, the expression of inflammatory chemokines and serum levels of EGFR ligands. A clinical trial was undertaken involving the escalation of cetuximab doses, guided by the grade of skin toxicity observed, such as no or low-grade, in metastatic colorectal cancer(the EVEREST study). The dose escalation of cetuximab was confirmed by a safety profile and had the tendency to achieve a higher response rate in KRAS wild-type patients. A large, prospective randomized trial is now ongoing(EVEREST 2) and the results of this trial may contribute to personalized medicine in KRAS wild-type colorectal cancer patients.展开更多
AIM To investigate biological mechanisms underlyingpyruvate kinase M2 isoform (PKM2) regulation of cellmigration and invasion in hepatocellular carcinomacells.METHODS: HepG2 and Huh-7 hepatocellularcarcinoma cell l...AIM To investigate biological mechanisms underlyingpyruvate kinase M2 isoform (PKM2) regulation of cellmigration and invasion in hepatocellular carcinomacells.METHODS: HepG2 and Huh-7 hepatocellularcarcinoma cell lines were stably transfected and culturedin DMEM (HyClone, Logan, UT, United States). Toinvestigate the effects of PKM2 on cellular proliferation,hepatocellular carcinoma cells were subjected tothe Cell Counting Kit-8 (Dojindo, Kamimashiki-gun,Kumamoto, Japan). And investigate the effects ofPKM2 on cell signal pathway related with migrationand invasion, Western immunoblotting were used tofind out the differential proteins. All the antibody usedwas purchaseed from Cell Signal Technology. In orderto explore cell motility used Transwell invasion andwound healing assays. The transwell plate with 0.5mg/mL collagen type Ⅰ (BD Bioscience, San Jose, CA)-coated filters. The wound-healing assay was performedin 6-well plates. Total RNA was extracted using TRIzolreagent (Invitrogen, CA, United States) and thenreverse transcription was conducted. Quantitativereverse transcription-polymerase chain reaction (PCR)analysis was performed with the ABI 7500 real-timePCR system (Applied Biosystems). We further usedigital gene expression tag profiling and identificationof differentially expressed genes.RESULTS: The cells seeded in four 96-well plates weremeasured OD450 by conducted Cell Counting Kit-8.From this conduction we observed that both HepG2and Huh-7 hepatocellular carcinoma cells with silencedPKM2 turn on a proliferate inhibition; however, cellmigration and invasion were enhanced compared withthe control upon stimulation with epidermal growthfactor (EGF). Our results indicate that the knockdownof PKM2 decreased the expression of E-cadherin and enhanced the activity of the EGF/EGFR signalingpathway, furthermore up-regulate the subsequentsignal molecular the PLCγ1 and extracellular signalregulatedkinase 1/2 expression in the hepatocellularcarcinoma cell lines HepG2 and Huh-7, which regulatescell motility. These variations we observed were due tothe activation of the transforming growth factor beta(TGFβ) signaling pathway after PKM2 knockdown. Wealso found that the expression of TGFBRI was increasedand the phosphorylation of Smad2 was enhanced.Taken together, our findings demonstrate that PKM2can regulate cell motility through the EGF/EGFR andTGFβ/TGFR signaling pathways in hepatocellularcarcinoma cells.CONCLUSION: PKM2 play different roles in modulatingthe proliferation and metastasis of hepatocellularcarcinoma cells, and this finding could help to guide thefuture targeted therapies.展开更多
探讨了0.4 m T工频磁场(Power frequency magnetic field,PFMF)暴露影响人羊膜成纤维(Human amniotic epithelial,FL)细胞迁移的分子机制。利用免疫荧光染色和western blotting方法检测了磁场暴露对FL细胞内黏着斑调控蛋白黏着斑激酶(Fo...探讨了0.4 m T工频磁场(Power frequency magnetic field,PFMF)暴露影响人羊膜成纤维(Human amniotic epithelial,FL)细胞迁移的分子机制。利用免疫荧光染色和western blotting方法检测了磁场暴露对FL细胞内黏着斑调控蛋白黏着斑激酶(Focal adhesion kinase,FAK)、桩蛋白(Paxillin)的分布、含量和部分关键活性蛋白的影响;通过细胞划痕实验,验证了磁场通过激活FAK从而激活细胞的迁移反应。结果表明:相较于对照组,PFMF暴露导致黏着斑总数量上升,并诱导信号蛋白FAK与桩蛋白更集中地分布在细胞边缘处,导致桩蛋白的总含量增加;FAK蛋白总含量无显著性变化,但其Y397磷酸化水平明显上调;用PD153035(PD)抑制表皮生长因子受体(Epidermal growth factor receptor,EGFR)的活性,或用FAK Inhibitor 14(FI)抑制Y397-FAK磷酸化,PFMF或表皮生长因子(Epidermal growth factor,EGF)引起的上述现象均消失;细胞划痕实验结果显示,PFMF促进了细胞的迁移,但当Y397-FAK被FI抑制后,PFMF对细胞迁移的促进效果几乎全部消失;PFMF引发的这些变化均与EGFR的配体EGF对细胞迁移产生的效果类似。0.4 m T工频磁场通过激活EGFR而激活黏着斑信号通路中的FAK、桩蛋白,调控黏着斑的形成与分布,在细胞前端协助片状伪足的延展,从而诱发细胞迁移。展开更多
4-Anilinoquinazoline analogues stand out among many kinds of small molecules that inhibit the tyrosine kinase activities of epidermal growth factor receptor (EGFR), thus serving as significant molecular targets for ...4-Anilinoquinazoline analogues stand out among many kinds of small molecules that inhibit the tyrosine kinase activities of epidermal growth factor receptor (EGFR), thus serving as significant molecular targets for anticancer drug design. Herein, a series of novel perfluorocarbon (PFC) modulated 4-anilinoquinazolines were designed and prepared straightforwardly by nucleophilic substitution reaction of various anilinoquinazolines and PFC-derived methanesulfonate. In the presence of base, the reaction proceeded smoothly to afford a wide range of 4-anilino- quinazolines with different substituents on aniline moiety in good to high yields. Furthermore, the PFC-modified analogues of gefitinib and erlotinib were also obtained in 93% and 90% respectively, which may have potential for developing new inhibitors of the epidermal growth factor receptor (EGFR) tyrosine kinase and fluorinated contrast agents (CA) for 19F MRI.展开更多
文摘1文献来源 Becker A, Crombag L, Heideman DA, et al. Retreatment with Erlotinib: Regain of TKI sensitivity following a drug holiday for patients with NSCLC who initially responded to EGFR-TKI treatment [J]. Eur J Cancer, 2011,47(17) :2603-2606.2
文摘目的:观察微小核糖核酸-134-5p(mi R-134-5p)转染对宫颈癌细胞增殖和凋亡的影响,验证其可能的分子机制。方法:收集湖北医药学院附属人民医院肿瘤中心2016年5月至8月收治的8名宫颈癌患者肿瘤组织和相应癌旁组织。利用lipofectamine 2000将mi R-134-5p mimics转染至宫颈癌Hela和Si Ha细胞。采用MTT法和集落形成实验检测细胞增殖活性;流式细胞术(FCM)检测细胞周期和细胞凋亡;q RT-PCR检测宫颈癌组织和细胞mi R-134-5p m RNA表达以及宫颈癌细胞EGFR m RNA表达;Western blotting检测宫颈癌细胞EGFR信号通路相关蛋白的表达。结果:宫颈癌组织mi R-134-5p m RNA表达显著低于癌旁组织(P<0.01)。和转染mi R-NC的Hela和Si Ha细胞比较,转染mi R-134-5pmimics的宫颈癌Hela和Si Ha细胞mi R-134-5p m RNA表达显著升高;细胞增殖能力显著降低(转染第5天,Hela细胞:1.06±0.13 vs 1.32±0.07;Si Ha细胞:1.12±0.10 vs 1.42±0.12,均P<0.05);形成的集落数减少;G0/G1期细胞比例显著上升,S期和G2/M期细胞比例显著下降;细胞凋亡率显著增加[Hela细胞:(26.53±13.48)%vs(3.25±1.74)%;Si Ha细胞:(30.49±12.04)%vs(5.10±2.86)%,均P<0.05];EGFR m RNA和EGFR蛋白表达显著下调,其中EGFR m RNA,Hela细胞下调58%(P<0.01),Si Ha细胞下调41%(P<0.05);EGFR下游靶蛋白p-AKT、p-ERK1/2和Cyclin D1蛋白及p EGFR蛋白表达显著下调。结论:mi R-134-5p可显著抑制宫颈癌细胞增殖并促进细胞凋亡,其可能的分子机制是通过抑制EGFR基因的表达,抑制EGFR通路的活化。
文摘Skin toxicity is a common symptom of anti-epidermal rowth factor receptor(EGFR) antibody treatment and is also a predictive marker of its efficacy in colorectal cancer patients. However, severe skin disorders induced by such antibodies negatively impact on the quality of life of patients and decreases drug compliance during treatment. If we can predict the high-risk group susceptible to severe skin toxicity before treatment, we can undertake the early management of any arising skin disorders and formulate a more accurate prognosis for anti-EGFR antibody treatment. Previous studies have identified molecular markers of skin toxicity induced by anti-EGFR antibody, such as EGFR polymorphisms, the expression of inflammatory chemokines and serum levels of EGFR ligands. A clinical trial was undertaken involving the escalation of cetuximab doses, guided by the grade of skin toxicity observed, such as no or low-grade, in metastatic colorectal cancer(the EVEREST study). The dose escalation of cetuximab was confirmed by a safety profile and had the tendency to achieve a higher response rate in KRAS wild-type patients. A large, prospective randomized trial is now ongoing(EVEREST 2) and the results of this trial may contribute to personalized medicine in KRAS wild-type colorectal cancer patients.
文摘AIM To investigate biological mechanisms underlyingpyruvate kinase M2 isoform (PKM2) regulation of cellmigration and invasion in hepatocellular carcinomacells.METHODS: HepG2 and Huh-7 hepatocellularcarcinoma cell lines were stably transfected and culturedin DMEM (HyClone, Logan, UT, United States). Toinvestigate the effects of PKM2 on cellular proliferation,hepatocellular carcinoma cells were subjected tothe Cell Counting Kit-8 (Dojindo, Kamimashiki-gun,Kumamoto, Japan). And investigate the effects ofPKM2 on cell signal pathway related with migrationand invasion, Western immunoblotting were used tofind out the differential proteins. All the antibody usedwas purchaseed from Cell Signal Technology. In orderto explore cell motility used Transwell invasion andwound healing assays. The transwell plate with 0.5mg/mL collagen type Ⅰ (BD Bioscience, San Jose, CA)-coated filters. The wound-healing assay was performedin 6-well plates. Total RNA was extracted using TRIzolreagent (Invitrogen, CA, United States) and thenreverse transcription was conducted. Quantitativereverse transcription-polymerase chain reaction (PCR)analysis was performed with the ABI 7500 real-timePCR system (Applied Biosystems). We further usedigital gene expression tag profiling and identificationof differentially expressed genes.RESULTS: The cells seeded in four 96-well plates weremeasured OD450 by conducted Cell Counting Kit-8.From this conduction we observed that both HepG2and Huh-7 hepatocellular carcinoma cells with silencedPKM2 turn on a proliferate inhibition; however, cellmigration and invasion were enhanced compared withthe control upon stimulation with epidermal growthfactor (EGF). Our results indicate that the knockdownof PKM2 decreased the expression of E-cadherin and enhanced the activity of the EGF/EGFR signalingpathway, furthermore up-regulate the subsequentsignal molecular the PLCγ1 and extracellular signalregulatedkinase 1/2 expression in the hepatocellularcarcinoma cell lines HepG2 and Huh-7, which regulatescell motility. These variations we observed were due tothe activation of the transforming growth factor beta(TGFβ) signaling pathway after PKM2 knockdown. Wealso found that the expression of TGFBRI was increasedand the phosphorylation of Smad2 was enhanced.Taken together, our findings demonstrate that PKM2can regulate cell motility through the EGF/EGFR andTGFβ/TGFR signaling pathways in hepatocellularcarcinoma cells.CONCLUSION: PKM2 play different roles in modulatingthe proliferation and metastasis of hepatocellularcarcinoma cells, and this finding could help to guide thefuture targeted therapies.
文摘探讨了0.4 m T工频磁场(Power frequency magnetic field,PFMF)暴露影响人羊膜成纤维(Human amniotic epithelial,FL)细胞迁移的分子机制。利用免疫荧光染色和western blotting方法检测了磁场暴露对FL细胞内黏着斑调控蛋白黏着斑激酶(Focal adhesion kinase,FAK)、桩蛋白(Paxillin)的分布、含量和部分关键活性蛋白的影响;通过细胞划痕实验,验证了磁场通过激活FAK从而激活细胞的迁移反应。结果表明:相较于对照组,PFMF暴露导致黏着斑总数量上升,并诱导信号蛋白FAK与桩蛋白更集中地分布在细胞边缘处,导致桩蛋白的总含量增加;FAK蛋白总含量无显著性变化,但其Y397磷酸化水平明显上调;用PD153035(PD)抑制表皮生长因子受体(Epidermal growth factor receptor,EGFR)的活性,或用FAK Inhibitor 14(FI)抑制Y397-FAK磷酸化,PFMF或表皮生长因子(Epidermal growth factor,EGF)引起的上述现象均消失;细胞划痕实验结果显示,PFMF促进了细胞的迁移,但当Y397-FAK被FI抑制后,PFMF对细胞迁移的促进效果几乎全部消失;PFMF引发的这些变化均与EGFR的配体EGF对细胞迁移产生的效果类似。0.4 m T工频磁场通过激活EGFR而激活黏着斑信号通路中的FAK、桩蛋白,调控黏着斑的形成与分布,在细胞前端协助片状伪足的延展,从而诱发细胞迁移。
文摘4-Anilinoquinazoline analogues stand out among many kinds of small molecules that inhibit the tyrosine kinase activities of epidermal growth factor receptor (EGFR), thus serving as significant molecular targets for anticancer drug design. Herein, a series of novel perfluorocarbon (PFC) modulated 4-anilinoquinazolines were designed and prepared straightforwardly by nucleophilic substitution reaction of various anilinoquinazolines and PFC-derived methanesulfonate. In the presence of base, the reaction proceeded smoothly to afford a wide range of 4-anilino- quinazolines with different substituents on aniline moiety in good to high yields. Furthermore, the PFC-modified analogues of gefitinib and erlotinib were also obtained in 93% and 90% respectively, which may have potential for developing new inhibitors of the epidermal growth factor receptor (EGFR) tyrosine kinase and fluorinated contrast agents (CA) for 19F MRI.