查找使用阿拉丁产品的科研文献

Synthesis of Novel Stigmasterol Derivatives with 1,2,3-Triazole Substituent at the C-3 Position and Their Cytotoxicity Toward HeLa, HGC-27, and HEK-293T Cells

Wang Yong,Wang Wei,Liu Cong-Jun,Gao Song,Xiong Yong-Ge,Gao Tian-Zeng,Li Wei-Shi,Li Jing-Jing

Eight new stigmasterol derivatives possessing 1,2,3-triazoles moiety were synthesized by a 1,3-dipolar cycloaddition reaction. All the compounds were characterized by IR, NMR, and HR-MS. In addition, human cervical cancer cells (HeLa), human gastric cancer cells (HGC-27), and human renal epithelial cells (HEK-293T) were selected to study the antitumor activity of the synthesized compounds. The results demonstrated that compounds 3c and 3d exhibited better inhibitory activity against the tested tumor cell lines. The IC 50 value of the inhibitory activity on HeLa was 52.3 μmol·L –1 and 31.4 μmol·L –1 , and 40.6 μmol·L –1 and 44.6 μmol·L –1 for HGC-27, respectively.

Cyclometalated Ru(II) β-carboline complexes induce cell cycle arrest and apoptosis in human HeLa cervical cancer cells via suppressing ERK and Akt signaling

Chen Jincan,Deng Yuanyuan,Wang Jie,Chen Suxiang,Peng Fa,He Xuerong,Liu Meijun,Luo Hui,Zhang Jingjing,Chen Lanmei

Abstract Two new cyclometalated Ru(II)-β-carboline complexes, [Ru(dmb) 2 (Cl-Ph-βC)](PF 6 ) (dmb = 4,4′-dimethyl-2,2′-bipyridine; Cl-Ph-βC = Cl-phenyl-9H-pyrido[3,4-b]indole; RuβC-3 ) and [Ru(bpy) 2 (Cl-Ph-βC)](PF 6 ) (bpy = 2,2′-bipyridine; RuβC-4 ) were synthesized and characterized. The Ru(II) complexes display high cytotoxicity against HeLa cells, the stabilized human cervical cancer cell, with IC 50 values of 3.2 ± 0.4 μM ( RuβC-3 ) and 4.1 ± 0.6 μM ( RuβC-4 ), which were considerably lower than that of non-cyclometalated Ru(II)-β-carboline complex [Ru(bpy) 2 (1-Py-βC)] (PF 6 ) 2 (61.2 ± 3.9 μM) by 19- and 15-folds, respectively. The mechanism studies indicated that both Ru(II) complexes could significantly inhibit HeLa cell migration and invasion, and effectively induce G0/G1 cell cycle arrest. The new Ru(II) complexes could also trigger apoptosis through activating caspase-3 and poly (ADP-ribose) polymerase (PARP), increasing the Bax/Bcl-2 ratio, enhancing reactive oxygen species (ROS) generation, decreasing mitochondrial membrane potential (MMP), and inducing cytochrome c release from mitochondria. Further research revealed that RuβC-3 could deactivate the ERK/Akt signaling pathway thus inhibiting HeLa cell invasion and migration, and inducing apoptosis. In addition, RuβC-3 -induced apoptosis in HeLa cells was closely associated with the increase of intracellular ROS levels, which may act as upstream factors to regulate ERK and Akt pathways. More importantly, RuβC-3 exhibited low toxicity on both normal BEAS-2B cells in vitro and zebrafish embryos in vivo. Consequently, the developed Ru(II) complexes have great potential on developing novel low-toxic anticancer drugs. Graphic Abstract

Betulinic acid induces apoptosis of HeLa cells via ROS-dependent ER stress and autophagy in vitro and in vivo

Chen Ping,Zhang Xueer,Fang Qiaomiao,Zhao Zhongxiang,Lin Chaozhan,Zhou Yuan,Liu Fangle,Zhu Chenchen,Wu Aizhi

Betulinic acid (BA), a naturally occurring lupane-type triterpenoid, possesses a wide range of potential activities against different types of cancer. However, the molecular mechanisms involved in anti-cervical cancer about BA were rarely investigated. Herein, the role of BA in cervical cancer suppression by ROS-mediated endoplasmic reticulum stress (ERS) and autophagy was deeply discussed. The findings revealed that BA activated Keap1/Nrf2 pathway and triggered mitochondria-dependent apoptosis due to ROS production. Furthermore, BA increased the intracellular Ca 2+ levels, inhibited the expression of Beclin1 and promoted the expression of GRP78, LC3-II, and p62 associated with ERS and autophagy. Besides, BA initiated the formation of autophagosomes and inhibited autophagic flux by the co-administration of BA with 3-methyladenine (3-MA) and chloroquine (CQ), respectively. The in vivo experiment manifested that hydroxychloroquine (HCQ) enhanced the apoptosis induced by BA. For the first time, we demonstrated that BA could initiate early autophagy, inhibit autophagy flux, and induce protective autophagy in HeLa cells. Thus, BA could be a potential chemotherapy drug for cervical cancer, and inhibition of autophagy could enhance the anti-tumor effect of BA. However, the interactions of signaling factors between ERS-mediated and autophagy-mediated apoptosis deserve further attention. Graphical abstract

A novel strategy for real-time and in situ detection of cytochrome c and caspase-9 in Hela cells during apoptosis

Qingqing Wen,Xi Zhang,Jiye Cai,Pei-Hui Yang

Cytochrome c (cyt c) and caspase-9 were critical biomarkers in mitochondria-mediated apoptosis. A novel electrochemical immunosensor was developed for in situ analysis of cyt c and caspase-9 in the cytosol. Gold nanoparticle–polydopamine (AuNP/PDA) composites were used to fabricate the interface of the sensor. The anti-cyt c or anti-caspase-9 functionalized-immunosensor provided a biomimetic interface for immunosensing of cyt c or caspase-9 in Hela cells during apoptosis. The changes in the expression level of cyt c and caspase-9 in the cytosol upon curcumin-induced apoptosis were detected by using the proposed method, and also the influence of different concentrations and incubation times of curcumin-induced Hela cells was investigated. This method achieved a linear range (0.1–100 μM) for standard cyt c and caspase-9, with a detection limit of 0.03 ± 0.01 μM for standard cyt c and 0.08 ± 0.02 μM for standard caspase-9. Moreover, this method was used to detect cells which could detect as low as 100 cells which expressed cyt c and caspase-9, and also the results are in good agreement with standard flow cytometry analysis. The developed electrochemical immunosensor offered a simple and rapid approach for sensitive evaluation of apoptosis markers with considerable specificity and reproducibility, and also the developed strategy could be of great importance in clinical diagnosis and therapeutic research.

The roles of breast cancer resistance protein (BCRP/ABCG2) and multidrug resistance-associated proteins (MRPs/ABCCs) in the excretion of cycloicaritin-3-O-glucoronide in UGT1A1-overexpressing HeLa cells

Shishi Li,Jinjin Xu,Zhihong Yao,Liufang Hu,Zifei Qin,Hao Gao,Kristopher W. Krausz,Frank J. Gonzalez,Xinsheng Yao

Cycloicaritin is a bioactive natural phenolic compound from Epimedium species. However, the glucuronidation and excretion which would influence oral bioavailability and pharmacokinetics of cycloicaritin still remain unknown. Here we aimed to establish UGT1A1 stably transfected HeLa cells, and to determine the contributions of BCRP and MRPs transporters to excretion of cycloicaritin-3- O -glucuronide. First, β-estradiol was used to validate the expression of active UGT1A1 protein in engineered HeLa1A1 cells. Furthermore, Ko143 (5 and 20 μM) led to a significant decrease (42.4%–63.8%, p  < 0.01) in CICT-3-G excretion and obvious accumulation (19.7%–54.2%, p  < 0.05) of intracellular CICT-3-G, while MK571 (5 and 20 μM) caused a significant reduction (46.8%–64.8%, p  < 0.05) in the excretion and obvious elevation (50.7%–85.2%, p  < 0.01) of intracellular level of CICT-3-G. Furthermore, BCRP knocked-down brought marked reduction in excretion rates of CICT-3-G (26.0%–42.2%, p  < 0.01), whereas MRP1 and MRP4-mediated silencing led to significant decrease in the excretion of CICT-3-G (23.8%–35.4%, p  < 0.05 for MRP1 and 11.9%–16.0%, p  < 0.05 for MRP4). By contrast, neither CICT-3-G excretion nor CICT-3-G accumulation altered in MRP3 knocked-down cells as compared to scramble cells. Taken together, BCRP, MRP1 and MRP4 were identified as the most important contributors for CICT-3-G excretion. Meanwhile, the UGT1A1 modified HeLa cells were a simple and practical tool to study UGT1A1-mediated glucuronidation and to characterize BCRP and MRPs-mediated glucuronide transport at a cellular level.

Activity Study of Cabotegravir Derivatives Inducing Mitochondrial Apoptosis in HeLa Cells

Song Yue,Zhan An,Wang Yimian,Zhang Xiaoting,Wang Lan,Hou Xixi

Cervical cancer ranks as the fourth most common cancer among women. Its progression is associated with accelerated cell cycle progression and inhibition of apoptosis. Given the unique potential of cabotegravir in suppressing cervical cancer cells and the widespread application of the 1,2,3-triazole scaffold in promoting mitochondrial apoptosis, we evaluated a series of cabotegravir derivatives incorporating 1,2,3-triazole for their anticancer potential. In vitro activity assays in HeLa cells revealed that compound 4v exhibited the most potent cytotoxicity (IC 50 = 3.06±0.17 μM). It significantly suppressed colony formation in a dose-dependent manner and induced marked morphological changes in HeLa cells. Further studies showed that compound 4v triggered apoptosis and caused G2/M cell cycle arrest. Western blot analysis demonstrated that 4v increased the Bax/Bcl-2 ratio, reduced levels of caspase-9, caspase-3, and PARP1, and promoted PARP1 cleavage, indicating that the mitochondrial apoptotic pathway was involved. In conclusion, compound 4v exhibited promising potential as a candidate for cervical cancer therapy.

Accurate identification of UDP-glucuronosyltransferase 1A1 (UGT1A1) inhibitors using UGT1A1-overexpressing HeLa cells

Hua Sun,Xiaotong Zhou,Baojian Wu

Electrochemical determination of caspase-3 using signal amplification by HeLa cells modified with silver nanoparticles

Wu Daohong,He Yuhan,Tong Liujuan,Wang Jianxiu,Liu Lin,Yi Xinyao,Hu Shengqiang

An electrochemical sensor capable of quantitative determination of caspase-3 activities was developed. A thiolated peptide whose sequence contained a caspase-3 cleaved site and a cell penetration sequence was preimmobilized onto an electrode. The quantification of caspase-3 was accomplished after cell penetration and the subsequent adsorption of silver nanoparticles (AgNPs). The oxidation current of AgNPs was found to be inversely proportional to the concentration of caspase-3 between 0.02 and 0.2 U/mL. A detection limit of 0.02 U/mL for caspase-3 was achieved due to the large number of positively charged AgNPs adsorbed onto the negatively charged cells. The proof of concept was demonstrated by monitoring the cleavage of surface-confined peptide substrates by caspase-3 in cell lysates. The current sensor could be extended to detect cells by replacing the surface-confined peptide with aptamers that recognize cells. Thus, the use of a cell as a matrix for AgNPs shows excellent potential for constructing electrochemical sensors and provides a useful alternative for sensor development in the future. Graphical abstract Cells modified with silver nanoparticles were utilized as the electrochemical readout of an electrochemical assay.

Titanium Dioxide Nanoparticles Induced HeLa Cell Necrosis under UVA Radiation through the ROS-mPTP Pathway

Runqing Geng,Yuanyuan Ren,Rong Rao,Xi Tan,Hong Zhou,Xiangliang Yang,Wei Liu,Qunwei Lu

Titanium dioxide nanoparticles (nano-TiO2), as a common nanomaterial, are widely used in water purification, paint, skincare and sunscreens. Its safety has always been a concern. Prior studies have shown that ultraviolet A (UVA) can exacerbate the toxicity of nano-TiO2, including inducing cell apoptosis, changing glycosylation levels, arresting cell cycle, inhibiting tumor cell and bacterial growth. However, whether the combination of UVA and nano-TiO2cause cell necrosis and its mechanism are still rarely reported. In this study, we investigated the cytotoxicity and phototoxicity of mixture crystalline nano-TiO2(25% rutile and 75% anatase, 21 nm) under UVA irradiation in HeLa cells. Our results showed that the abnormal membrane integrity and the ultrastructure of HeLa cells, together with the decreased viability induced by nano-TiO2under UVA irradiation, were due to cell necrosis rather than caspase-dependent apoptosis. Furthermore, nano-TiO2and UVA generated the reactive oxygen species (ROS) and caused the mitochondrial permeability transition pore (mPTP) of HeLa cells to abnormally open. Cell viability was significantly increased after adding vitamin C (VC) or cyclosporin A (CsA) individually to inhibit ROS and mPTP. Clearance of ROS could not only impede the opening of mPTP but also reduce the rate of cell necrosis. The results suggest the possible mechanism of HeLa cell necrosis caused by nano-TiO2under UVA irradiation through the ROS-mPTP pathway.Keywords:nano-TiO2;UVA;cell necrosis;reactive oxygen species;mitochondrial permeability transition pore

Imaging of lysosomal oxidative stress during autophagy with a ratiometric probe featuring a large probe-product spectral separation

Yinghua Guo,Xinru Wang,Yan Feng,Xiao Luo,Xiangming Meng,Xuhong Qian,Youjun Yang

Autophagy is a physiological process governing proteostasis and implicated in many diseases. Enhanced ROS levels was known to be associated with lysosomal autophagy and therefore a lyso-specific ROS probe could find applications in autophagy related studies. Following the probe-design principle of bridging-group modification, we previously developed a ROS probe capable of detect a broader range of reactive species than the existing probes. Additional merits of the probe include ratiometry-capability, minimal probe-product spectral crosstalk, resistance toward autoxidation at lysosomal acidity, and structural simplicity. It is the aim of this manuscript to render this probe lyso-specificity and examine its potentials to monitor autophagy-related ROS generation in HeLa cells treated by H 2 O 2 or 6-OHDA.

Characterization of Chrysin Glucuronidation in UGT1A1-Overexpressing HeLa Cells: Elucidating the Transporters Responsible for Efflux of Glucuronide

Enxi Quan,Huailing Wang,Dong Dong,Xingwang Zhang,Baojian Wu

Active transport of glucuronide out of cells is a critical process in elimination of drugs via the glucuronidation pathway. Here, HeLa cells were stably transfected with UGT1A1 and the contributions of BCRP and MRP family transporters to the cellular efflux of chrysin glucuronide (CG) were determined. The cDNA of UGT1A1 was introduced into HeLa cells using the lentiviral transfection method. The modified cells were functional in generation of the glucuronide from chrysin. Ko143 at 10–20 μM (a dual inhibitor of BCRP and UGT1A1) caused a marked decrease (51.3%–59.7%, P < 0.01) in the excretion rate and efflux clearance of CG. Likewise, MK-571 at 5–20 μM (an inhibitor of MRPs but an activator of UGT1A1) resulted in a significant reduction in the excretion rate (18.2%–64.0%, P < 0.01) and efflux clearance (37.0%–90.2%, P < 0.001). By contrast, dipyridamole and leukotriene C4 showed no inhibitory effects on CG excretion. The chemical inhibition indicated that excretion of CG was contributed by the MRP family transporters, whereas the role of BCRP was unclear. Furthermore, short hairpin RNA–mediated silencing of a target transporter led to a marked reduction in the excretion rate of CG (38.6% for BCRP, 39.3% for MRP1, 36.4% for MRP3, and 28.7% for MRP4; P < 0.01). Transporter silencing also led to substantial decreases in the efflux clearance (44.7% for BCRP, 60.4% for MRP1, 36.7% for MRP3, and 28.7% for MRP4; P < 0.01). The gene silencing results suggested that BCRP, MRP1, MRP3, and MRP4 were significant contributors to excretion of CG.

Acetaldehyde-modified-cystine as an enhanced fluorescent probe for intracellular glutathione imaging

Linlin Hu,Xing Wei,Jie Meng,Xiaoyan Wang,Xuwei Chen,Jianhua Wang

The development of fluorescent probes for detection of glutathione (GSH) has attracted extensive attentions due to its close association with cellular functions and diseases. Herein, a fluorescence enhancement sensing system based on acetaldehyde-modified-cystine (AMC) is reported for the detection of thiols, providing detection limits of 36 and 52 μmol L −1 for GSH and cysteine (Cys) respectively. Importantly, the fluorescence response of AMC is highly selective toward thiols with respect to the coexisting species in real biological matrixes. Considering the fact that GSH levels in cancer cells is significantly higher than those of Cys and other reducing species, the fluorescence enhancing response of AMC in cancer cells is dominated by GSH. Therefore, this provides the possibility for selective intracellular GSH imaging in living cancer cells, i.e., HeLa cells in the present study.