Cell Inhibition Rate Research Articles

NOTA analogue: A first dithiocarbamate inhibitor of metallo-β-lactamases

The emergence of antibiotic drug (like carbapenem) resistance is being a global crisis. Among those resistance factors of the β-lactam antibiotics, the metallo-β-lactamases (MBLs) is one of the most important reasons. In this paper, a series of cyclic dithiocarbamate compounds were synthesized and their inhibition activities against MBLs were initially tested combined with meropenem (MEM) by in vitro antibacterial efficacy tests. Sodium 1,4,7-triazonane-1,4,7-tris(carboxylodithioate) (compound 5) was identified as the most active molecule to restore the activity of MEM. Further anti-bacterial effectiveness assessment, compound 5 restored the activity of MEM against Escherichia coli, Citrobacter freundii, Proteus mirabilis and Klebsiella pneumonia, which carried resistance genes of blaNDM-1. The compound 5 was non-hemolytic, even at a concentration of 1000 µg/mL. This compound was low toxic toward mammalian cells, which was confirmed by fluorescence microscopy image and the inhibition rate of HeLa cells. The Ki value of compounds 5 against NDM-1 MBL was 5.63 ± 1.27 μM. Zinc ion sensitivity experiments showed that the inhibitory effect of compound 5 as a MBLs inhibitor was influenced by zinc ion. The results of the bactericidal kinetics displayed that compound 5 as an adjuvant assisted MEM to kill all bacteria. These data validated that this NOTA dithiocarbamate analogue is a good inhibitor of MBLs.

Synthesis, structures, fluorescence studies and cytotoxicity of a new Manganese(II) complex

ABSTRACTThe novel metal coordination polymer [Mn(L)2(H2O)2]n, was synthesized and characterized. Fluorescence and UV absorption spectroscopy indicates that the ligand and Mn(II) comploud can bind to fish sperm DNA (FS-DNA). Gel electrophoresis assay demonstrated the ability of the ligand and Mn(II) complex to cleave the HL-60 plasmid DNA. Apoptotic study shows the complexes exhibited significant cancer cell(JEKO-1 and KB) inhibitory rate. The results indicate that the complexe and the ligand bound to DNA with different binding affinity, in the order the Mn(II) complex > the ligand. Oxygen and nitrogen atoms which chelated with Mn(II) forming hydrogen bonds with N6 atom of adenine and O4 atom of thymine to affect the stability of DNA from Molecular docking study.

Scutellarin Inhibits Proliferation, Invasion, and Tumorigenicity in Human Breast Cancer Cells by Regulating HIPPO-YAP Signaling Pathway.

BackgroundThe present study aimed to investigate the effects of Scutellarin on proliferation, invasion and tumorigenicity in human breast carcinoma MCF-7 cells and its associated molecular mechanisms.Material/MethodsThe MCF-7 cells were cultured with varies of concentrations of Scutellarin in vitro. The proliferation, invasion, and apoptosis of MCF-7 cells were studied via CCK-8 assay, transwell assay, and flow cytometry. In vivo expression of the HIPPO pathway key proteins YAP and p-YAP of MCF-7 cells were analyzed by immunohistochemistry.ResultsThe inhibition rates of Scutellarin-treated MCF-7 cells were 40.1%, 58.7%, and 70.6% for 24, 48, and 72 h, respectively. The MCF-7 cell proliferation was significantly inhibited by Scutellarin. Treating MCF-7 cells with Scutellarin led to invasion inhibition. The rates apoptotic cells were between 12.4±1.9% and 23.9±2.1% in 40–120 μM Scutellarin-administrated groups, which had a significant rise compared with the control group (7.8±1.9%, P<0.05). Scutellarin significantly inhibited MCF-7 xenograft tumor growth. Immunohistochemical analysis showed that the inhibition of tumor growth in Scutellarin-treated mice was associated with increased p-YAP and decreased YAP expression in vivo.ConclusionsScutellarin-treated breast carcinoma MCF-7 cells had significantly inhibited growth and induced apoptosis, which is associated with induction of autophagy through regulation of the HIPPO-YAP signaling pathway, providing support to the clinical use of Scutellarin-based medication to achieve optimized outcome in patients with breast carcinoma.

Fructose‑1,6‑bisphosphatase‑1 decrease may promote carcinogenesis and chemoresistance in cervical cancer.

Fructose-1,6-bisphosphatase-1 (FBP1), a gluconeogenesis rate-limiting enzyme expressed in various tissues, is important in the carcinogenesis of various cancers. To evaluate the association of FBP1 expression and carcinogenesis and chemoresistance in cervical cancer, the present study analyzed 140patients of squamous cell carcinoma of cervical cancer (CSCC) who had adjuvant concurrent chemoradiation therapy following radical surgery. By detecting FBP1 protein expression in paraffin‑embedded tumor tissues through immunohistochemistry, it was observed that 50% of the cases had a low expression of FBP1, which was associated with a shorter overall survival time (P=0.011). In addition, FBP1 mRNA level was downregulated in tumor tissues, compared with cervical normal tissues. Among the tumor‑associated prognostic factors, loss of FBP1 expression (χ2‑test, P=0.025) was significantly associated with the tumor recurrence and greater tumor stage of cervical cancer patients (2‑test, P<0.0001). In 3‑(4,5)‑dimethylthiahiazo(‑z‑y1)-3,5-diphenytetrazoliumromide (MTT) assay of primary tumor cells, the median invitro inhibition rate of cisplatin, carboplatin, nedaplatin, and oxaliplatin was 62, 47, 58 and 52%, respectively. Although there was no significant association between FBP1 expression and invitro tumor inhibition rates of primary tumor cells, overexpression of FBP1 markedly suppressed carcinogenesis and restored the chemosensitivity to cisplatin in cervical cancer cell lines of HeLa and CaSki. Overall, decreased levels of FBP1 may be used as a predictor for poor prognosis of cervical cancer patients, however the mechanism requires further investigation.

Effects of calcium channel on ovarian cancer cells.

The aim of the present study was to examine the effects of calcium channel protein on ovarian cancer cells. The expression of calcium channel protein in normal ovarian cells and ovarian cancer cells was detected by fluorescence quantitative PCR. Subsequently, the ovarian cancer cells were added to calcium channel protein activator media at various concentrations of 0, 1, 4, 8, 12, 16 and 20 mmol/l. The concentration of calcium ion in different samples was produced, and using an MTT assay, ovarian cancer cell activity in various samples was detected. Finally, a flow cytometer was used to explore the apoptosis rate. It was found that there was a significant difference between the expression of calcium channel protein in normal ovarian tissue and ovarian cancer cells (P<0.05), as well as a significant difference of calcium concentration among various samples (P<0.05). When the concentrations of calcium channel activator were 1, 4, 8, 12, 16 and 20 mmol/l, the values of the ovarian cancer cell inhibition rates were 4.6, 21.3, 48.3, 67.9, 52.8 and 31.8%, respectively. It showed that the calcium channel activator inhibited the proliferation of ovarian cancer cells to a certain extent, in a dose-dependent manner, especially when the concentration was at 12 mmol/l at which the intracellular calcium concentration was similar to that in normal ovarian cells. In conclusion, calcium ions play an important role in promoting cell proliferation of ovarian cancer cells, and they were involved in apoptosis of ovarian cancer cells to some extent, which regulates apoptosis by controlling the content of intracellular calcium.

Effects of ganglioside GM3 on proliferation and cell cycle of multiple myeloma cell line U266

Objective To observe the effect of ganglioside GM3 on the proliferation and cell cycle of multiple myeloma (MM) cell line U266. Methods The experimental groups were treated with 20, 40, 80, 160 μmol/L GM3, while the control group was not given GM3. The growth inhibition effect of GM3 on U266 cells after 48 hours were measured by using methyl thiazolyl tetrazolium (MTT) method. The cell cycle was tested by flow cytometry with PI labeling. Results The results indicated that the GM3 displayed anti-proliferative effects on U266 cells in a dose-dependent manner. The cell inhibition rates were (13±4)%, (26±4)%, (47±6)%, (55±10)% in different dose of GM3 (20, 40, 80, 160 μmol/L), respectively. There was a difference between the experimental group and the control group (F= 93.063, P < 0.05). At the same time, the cell cycle analysis results showed that the U266 cell ratio in S phase was increased from (22.6±3.7)% to (71.5±3.8)% (P < 0.01) and in G2/M phase was decreased from (42.6±2.5)% to (0.8±0.6)% (P < 0.05) while in G0/G1 phase was not significantly changed. Conclusion GM3 can inhibit the proliferation of MM cell line U266 by blocking them in the S phase. Key words: Gangliosides; Multiple myeloma; Cell proliferation; Cell cycle

Bortezomib could down-regulate the expression of RANKL, inhibit cell proliferation and induce cell apoptosis in the human myeloma cell line RPMI 8226 by activating casepase-3.

In spite of bortezomib being developed and demonstrated as a safe drug therapy for multiple myeloma (MM), the role of bortezomib-induced receptor activator of nuclear factor (NF)-κB ligand (RANKL) in the MM cell lines remains to be understood. Thus the present study aims to explore the impact of bortezomib on RANKL expression, cell growth and apoptosis in human myeloma cell line RPMI 8226. Four experiment groups were set according to different concentrations of bortezomib, namely blank group (treated with DMEM solution free of other drugs), low-dose group (treated with 10 nmol/L bortezomib), middle-dose group (treated with 20 nmol/L bortezomib) and high-dose group (treated with 40 nmol/L bortezomib). Western blotting was adopted to detect RANKL protein expression. MTT assay was performed to detect cell proliferation. Flow cytometry was used to analyze cell cycle and apoptosis. Spectrophotometry was applied to determine caspases-3 activity. Compared with the blank group, the RANKL protein expression, cell number at the S stage was reduced while cell inhibition rate, cell apoptosis rate and caspase-3 activity enhanced remarkably in the low-dose, middle-dose and high-dose groups with dose-dependent manner. Compared with those treated with bortezomib (20 nmol/L and 40 nmol/L) for 6 h, the RANKL expression was down-regulated, cell inhibition rate was increased, cells at the S stage were reduced, cell apoptosis rate was enhanced, and caspase-3 activity elevated in the RPMI 8226 cells as treated with bortezomib for 24 h, with a dose- and time-dependent manner. Bortezomib could reduce the RANKL expression, inhibit cell proliferation and activate caspase-3 activity to induce cell apoptosis in RPMI 8266 cells.

Tetrandrine and arsenic trioxide synergistically inhibit proliferation of HCC1937 triple negative breast cancer cells

ObjectiveTo evaluate the effects of tetrandrine plus arsenic trioxide on HCC1937 cells, a triple negative breast cancer cell line, and to explore possible mechanisms. MethodsThe 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide method was used to compare the antiproliferative effects of tetrandrine, arsenic trioxide alone and tetrandrine plus arsenic trioxide on HCC1937 cells. The median-effect principle (Chou-Talalay combination index method) was used to examine the interaction between the two drugs. Flow cytometry was used to evaluate effects of treatment with tetrandrine, arsenic trioxide or the combination of both on HCC1937 cell apoptosis. Real-time polymerase chain reaction and western blotting were performed to evaluate changes in apoptosis-related gene expression and protein levels. ResultsTetrandrine and arsenic trioxide each inhibited HCC1937 cell proliferation in a dose-dependent manner. The cell inhibition rate of HCC1937 cells treated with a combination of tetrandrine and arsenic trioxide was significantly higher than with either compound alone. The two drugs produced a synergistic effect when the inhibition rate was 20%-40%. Flow cytometry results showed that treatment with the two drugs increased the proportion of apoptotic cells. In the combination treated group, caspase-3 activation and PARP cleavage were significantly higher than in the other groups. Moreover, Bcl-2 and survivin expression were decreased, whereas that of both Bid and Bax was increased. ConclusionThese findings demonstrated that tetrandrine plus arsenic trioxide had synergistic efficacy on induction of apoptosis in HCC1937 cells.

Effects of bufalin combined with doxorubicin on the proliferation and apoptosis of human lung cancer cell line A549 in vitro

To explore the effects of bufalin (BUF) combined with doxorubicin (DOX) on the proliferation and apoptosis in human lung cancer cell line A549 in vitro. Methods: Methyl thiazolyl tetrazolium (MTT) assay was used to measure the inhibitory effects of BUF, DOX and their combination on the growth of A549 cells. Hoechst 33342 staining was used to observe the changes of nucleus. Flow cytometry was used to investigate the apoptosis and cell cycle distribution of A549 cells. Western blot was used to examine the expression of apoptotic protein. Results: BUF and DOX showed inhibitory effect on the A549 cells in a dose and time-dependent manner. Compared with BUF or DOX alone, combination of BUF (1, 20, 100 nmol/L) with DOX (1.0 μg/mL) could significantly increase the growth inhibition rate of A549 cells at 24, 36, 72 h, respectively (all P<0.05). BUF and DOX alone could induce apoptosis, and their combination could significantly increase the apoptosis ratio. In addition, BUF combined with DOX could block the cell stage of A549 cells, keep the cell stage stay in S stage and up-regulate the expression of caspase-3. Conclusion: BUF combined with DOX can significantly inhibit the proliferation of A549 cells, which might be related to the induction of apoptosis, cell cycle S phase arrest and caspase-3 up-regulation.

Clinical significance of σ1 receptor over-expression in cervical cancer and the effect of its synthetic ligands on the growth of cervical cancer cells

Objective: To explore the role of σ1 receptor (σ1R) in the clinical prognosis of cervical cancer,and provide a theoretical basis for σ1R targeted molecular therapy through observing the inhibition of synthetic σ1R-specific ligand compounds on the growth of cervical cancer cells. Methods: (1) Immunohistochemical or immunocytochemistry staining were respectively used to detect the expression and localization of σ1R protein. (2) The Cancer Genome Atlas (TCGA) data set was used to validate our results. (3) Two series of 4 novel σ1R ligand compounds were synthesized by altering the N-terminal substituents on the piperidine ring of the prezamicol analogue, named as 14a, 14e, 15c and 15f. Methyl thiazolyl-tetrazolium (MTT) assay was detect the anti-proliferative effect of the four compounds on HeLa and SiHa cells. Compound 14a with potent inhibitory activity and the highest specificity of σ1R was selected for further experiments. Scratch test was observed the migration effect of compound 14a on HeLa and SiHa cells. Flow cytometry was determined cell cycles and apoptosis. Results: (1) Immunostaining of σ1R protein was located in the cytoplasm and nucleus of cervical epithelium. The expression of cervical squamous cell carcinoma (SCC) was significantly higher than those of high-grade squamous intraepithelial lesion (HSIL) or normal cervical tissues. There was no significant difference in the expression of σ1R between HSIL and normal cervical tissues. σ1R expression in cervical adenocarcinoma (AC) was higher than that in SCC (P=0.020). The nuclear expression rate of σ1R in AC (10/18) was higher than that of SCC (27.1%, 19/70; P=0.024). The median overall survival (MOS) of σ1R-positive SCC patients was lower than that of σ1R-negative patients [(45.8±3.1) vs (51.7±2.9) months, P=0.045]. MOS of the patients with σ1R nuclear positive SCC was lower than that of non-nuclear staining [(38.9±3.8) vs (48.7±2.1) months, P=0.022]. MOS of the patients with σ1R nuclear positive AC was lower than that of non-nuclear staining [(35.0±6.3) vs (44.2±4.2) months, P=0.034]. (2) Analysis of TCGA data showed that σ1R expression of in SCC was correlated with age (P=0.005). σ1R expression in AC was significantly associated with advanced stage, lymphnode metastasis and vascular invasion (all P<0.05). MOS of AC patients with σ1R overexpression was significantly lower than that of the patients with low expression (P=0.034). There was no significant difference in the MOS of different expression of σ1R mRNA in SCC patients(P=0.930). (3) MTT assay showed that these four compounds could suppressed the growth of HeLa and SiHa cells in time- and dose-dependent manner. The growth inhibition rates of HeLa and SiHa cells at 48 hours treated by combination of different concentrations of nedaplatin (NDP) with compound 14a (6 μmol/L) were significantly higher than those treated by NDP alone. Compound 14a (30 μmol/L) significantly inhibited the migration (both P<0.01) and induced the apoptosis of HeLa or SiHa cells (both P<0.01). Conclusions: σ1R is over-expressed in cervical cancer and HSIL. σ1R nuclear expression is an important marker of AC. σ1R over-expression, especially σ1R nuclear expression is associated with the poor prognosis of cervical cancer. Our study is mostly consistent with cervical cancer data of TCGA. These results suggest that the novel synthetic prezamicol analogues 14a for σ1R could inhibit the growth of cervical cancer cells and cell migration through inducing apoptosis and arresting cell cycle in G(0)/G(1) period, enhance NDP-induced cytotoxicity.

Effect of Rat C6 Glioma Cells in Varying Concentrations of Cultured in Isorhamnetin

Objective: To investigate the effects of different concentrations of isorhamnetin on C6 rat glioma cells in vitro from January 2015 to June 2015. Methods: The blank control group, blank solvent control group and four concentration groups were used to observe the cell growth status under a microscope. MTT colorimetric assay was used to detect the effect of isorhamnetin on C6 glioma cells in vitro and the cell inhibition rate And survival rate were measured. The apoptotic and apoptotic rates were measured by flow cytometry in the treatment group and the control group. The relationship between the different concentrations of isorhamnetin and C6 glioma cell apoptosis was analyzed the total protein was extracted and the total AKT protein and Ser473 AKT protein content were detected by Western blotting. The rat model of glioma was constructed by SD rats.Five days of isorhamnetin was continuously fed and the plasma was detected by high-performance liquid chromatography,liver, brain tissue isorhamnetin content.Â

The effect of Yiqi-jianpi-jiedu recipe decoction containing serum on oxaliplatin-induced HepG2 human hepatoma cell

Objective To explore the effect of Yiqi-jianpi-jiedu recipe decoction containing serum on Oxaliplatin-induced HepG2 Human hepatoma cell survivin, caspase-3, caspase-9 expression. Methods The serum pharmacological method was used to prepare nourishing Qi and invigorating spleen decoction containing serum of rat. The HepG2 human hepatoma cells were divided into Chinese herb containing serum group, chemotherapy with Oxaliplatin (OXA) group and the combination group (medicine containing serum and OXA), and the blank group without drugs. Apply MTT assay, Western Blotting method and RT-PCR method to detect the cell inhibition rate and survivin, caspase-3, caspase-9 protein and gene expression changes. Results Compared with the blank group, the inhibition rate of HepG2 Human hepatoma cell was significantly higher in Chinese herb containing serum group, chemotherapy group and the combination group (P < 0.05). Compared with thechemotherapy group and Chinese herb containing serum group, the Survivin (0.151 ± 0.054 vs. 0.288 ± 0.089, 0.375 ± 0.063) and the expression of Survivin mRNA (0.205 ± 0.091 vs. 0.487 ± 0.073, 0.725 ± 0.092) of the conbination group downgraded significantly lower (P < 0.05 or P< 0.01). Compared with chemotherapy group and Chinese herb containing serum group, the caspase-3 (0.821 ± 0.079 vs. 0.634 ± 0.098, 0.487 ± 0.102), caspase-9 (0.901 ± 0.047 vs. 0.709 ± 0.054, 0.402 ± 0.012) expression, caspase-3 mRNA (1.928 ± 0.226 vs. 1.564 ± 0.195, 1.287 ± 0.312) and caspase-9 mRNA (2.063 ± 0.517 vs. 1.536 ± 0.084, 1.019 ± 0.182) expression of the conbination group upgraded significantly higher (P < 0.05 or P < 0.01). Conclusions Nourishing-Qi and invigorating spleen decoction containing serum may affect Oxaliplatin-induced HepG2 apoptosis mechanism through increasing the suppression on Survivin and promoting the expression of caspase-3 and caspase-9 protein and genes. This research provided important experimental basis for clinical application of traditional Chinese medicine treating liver cancer. Key words: Apoptosis; Seropharmacology (TCD); Yiqi-jianpi-jiedu recipe

MiR-33b-5p sensitizes gastric cancer cells to chemotherapy drugs via inhibiting HMGA2 expression

MicroRNAs (miRNAs) are internal, non-coding, and ∼22 nt small RNAs that display cell- and tissue-specific expression. They play important regulatory roles in cell proliferation and chemo-sensitivity. This study focused on tumor-suppressive miR-33b-5p expression as well as its role in gastric cancer. MiR-33b-5p was found low expression in gastric cancer cell lines. Functionally, western blots and the luciferase reporter assay were used to confirm that HMGA2 was the potential target of miR-33b-5p. Next, we used CCK-8 kits to analyze the effect of miR-33b-5p combined chemotherapy drugs on cell inhibition rate, and flow cytometry to analyze cells apoptosis. Colony formation ability was determined by plating at 500 cells per well into six-well plates and culturing for 15 d. The results showed that upregulation of miR-33b-5p decreased expression of HMGA2 and inhibited gastric cancer cell growth as well as sensitized gastric cancer cells to chemotherapy drugs. MiR-33b-5p overexpression hindered luciferase activity of HMGA2,3′-untranslated region-based reporter construct in 293 T cells. These data demonstrate that miR-33b-5p may be a potential therapeutic target for gastric cancer and function as tumor-suppressive miRNA through targeting HMGA2 in gastric cancer.

Exosome-transmitted microRNA-19a promotes drug resistance in breastcancer by targeting phosphatase and tensin homolog/protein kinase B/p53 axis

Objective To investigate the specific effect and underlying molecular mechansim of microRNA (miRNA, miR)-19a through MCF-7/A secrete dexosomes in drug resistance transfer. Methods Exosomes were extracted from the supernatant of cultured MCF-7/A cells by fractionation ultracentrifugation and were identified by transmission electron microscopy and Nanosight. GFP-MCF-7/S, a breast cancer sensitive cell line stably expressing green fluorescent protein (GFP), was constructed by recombinant lentiviral vector with GFP. The co-cultured experiments of cell-cell and cell-exosomewere designed to observe thephenomenon of drug resistance transfer. Real-time quantitative polymerase chain reaction(Real-time PCR), Western blotting and cell transfection techniquewere used to determine the specific effects of miR-19a and phosphatase and tensin homologue deleted on chromosometen (PTEN)/protein kinase B (Akt)/p53 axis in the progress of drug resistance. Results Compared to MCF-7/S, the expression of miR-19a was significantly increased in MCF-7/A cells (1.03±0.16 vs. 3.56±0.58, P=0.002), down-regulaion of miR-19a apparently increased sensitivity to Adriamycin (ADM) in MCF-7/A cells [(41.17±10.65)% vs. (23.45±8.89)%, P=0.011]. Transmission electron microscopy showed that the exosomes cells had adiameter of 40-100 nm and exhibited round or elliptic shape secreted by both two type cancer cells. The cell inhibition ratio of ADM detected by the intensity of GFP and cell counting kit-8 (CCK-8) assay showed that cell inhibition rate of MCF-7/S+ MCF-7/A and MCF-7/S+ MCF-7/A exos group were significantly lower than the MCF-7/S+ MCF-7/S [(31.50±6.68)% vs. (51.20±12.44)%, P=0.006] and MCF-7/S+ MCF-7/S exos group [(27.67±6.02)% vs. (49.65±10.52)%, P=0.000], which companied with the increasing expression miR-19a (1.01±0.13 vs. 2.67±0.62, P=0.000). Western blotting assay showed that miR-19 in exosomes secreted by drug resistant cancer cells significantly inhibited the expression of PTEN (1.15±0.18 vs. 0.67±0.13, P=0.000) and p53 protein (0.99±0.18 vs. 0.49±0.11, P=0.005), whereas increases phosphorylation of Akt (1.15±0.18 vs. 0.67±0.13, P=0.000), which promoted the occurrence of drug resistance in receptor cells. Conclusion MiR-19a transfers drug resistance between breast cancer cells by exosome secreted from drug-resistance cancer cells through the regulation of PTEN/Akt/p53 axis in receptor cancer cells. Key words: Exosome; MicroRNA-19a; Breast cancer; Drug assistant; Phosphatase and tensin homologue deleted on chromosometen/protein kinase B/p53 axis

Two novel dinuclear ellipsoid Ni(II) and Co(II) complexes bridged by 4,5-bis(pyrazol-1-yl)phthalic acid: Synthesis, structural characterization and biological evaluation

Two novel complexes, [Ni2(L)2(H2O)3]·4(H2O) (1) and [Co2(L)2(H2O)3]·5(H2O) (2) [H2L = 4,5-bis(pyrazol-1-yl) phthalic acid] were synthesized and characterized by spectroscopy (IR, 1H NMR), and elemental analysis. The structures for the complexes were determined by X-ray crystallography providing the dinuclear ellipsoid Ni(II) and Co(II) complexes bridged by 4,5-bis(pyrazol-1-yl)phthalic acid ligands with same coordination modes. The interaction capacity of the complexes with FS-DNA (fish sperm DNA) has been investigated by UV and fluorescence spectroscopy. Gel electrophoresis assay demonstrated the ability of the complexes to cleave the pBR322 plasmid DNA. The cytotoxic activity of the complexes was tested against two different cancer cell lines, HeLa (human cervix epithelia carcinoma cells) and KB (human oral epithelial carcinoma cells), exhibiting significant cancer cell inhibitory rate. Furthermore, flow cytometry experiments and morphological apoptosis studies showed that the complexes induced apoptosis of KB tumor cell lines. The good visualization images supported with the experimental results of structure-activity relationship.

Synthesis and effect on SMMC-7721 cells of new benzo[c, d]indole rhodanine complex merocyanines as PDT photosensitizers

Photodynamic therapy (PDT) represents a modern and noninvasive therapeutic approach, however, it relies on the development of photosensitizers. Here five new benzo[c,d]indole rhodamine complex merocyanines (BIRCM) D1-D5, displaying low dark toxicity and significant photo toxicity, were synthesized as PDT photosensitizers, and characterized by 1H NMR, IR, UV–Vis and HRMS. The investigation of their absorption spectra in different solvents showed that the absorption maxima and molar extinction coefficient were in the region 507–679 nm and 0.21 × 104–1.27 × 105 L · mol−1cm−1, respectively. The evaluation of PDT activity showed that only irradiation could not kill SMMC-7721 cells, and the cell survival rate and inhibition rate at the application dose and duration was 92%–87% and 78%–49%, respectively. Especially, using D2, absorbed in the red zone, as photosensitizer for PDT analyzed its effect on SMMC-7721 cells survival, it could be found that the cell survival rate was 92% without irradiating and the cell inhibited rate was 78% under irradiating at concentrations of 2.5 × 10−6 mol/L, displaying low dark toxicity and high photo toxicity, which was valuable for PDT of some microvascular diseases or other superficial diseases.

The role of FOXO3a-Bim signaling in triptolide induced bladder cancer T24 cells apoptosis

Objective: To investigate the role of FOXO3a-Bim signaling in triptolide induced bladder cancer T24 cells apoptosis. Methods: T24 cells were used and divided into control group, triptolide group(50 nmol/L), MK2206 group(50 nmol/L triptolide+ 5 μmol/L MK2206), FOXO3a-siRNA group(50 nmol/L triptolide+ 100 nmol/L FOXO3a-siRNA), Bim-siRNA group (50 nmol/L triptolide+ 100 nmol/L Bim-siRNA). MTT assay was used to analyze the cells growth inhibition.Annexin V/PI staining was implemented to detect cell apoptosis rate, the expression of p-Akt, Akt, p-FOXO3a, FOXO3a, Bim, Bax.Cleaved-caspase 3 was analyzed by Western blot. Results: After treatment with triptolide 25, 50, 100, 250 nmol/L, the cell growth inhibition rates at 24 hours(17%±9%, 24%±5%, 43%±8%, 61%±8%), 48 hours (20%±7%, 34%±6%, 56%±7%, 74%±5%) and 72 hours(32%±8%, 41%±7%, 69%±7%, 84%±3%) were significantly higher than control group respectively.The IC(50) at 24, 48, 72 hours were (113±10), (91±8), (68±5) nmol/L; the cell apoptosis rates at 24 hours (10%±4%, 15%±5%, 29%±8%, 46%±8%), 48 hours (16%±5%, 24%±6%, 40%±7%, 55%±9%) and 72 hours (27%±4%, 38%±5%, 50%±9%, 65%±8%) were significantly increased (P<0.05). Western blot showed that triptolide reduced the expression of p-Akt, p-FOXO3a and increased the expression of Bim, Bax, cleaved-caspase 3.The cell inhibition rate in Triptolide group (30%±8%) was significantly higher than that in the control group (P<0.05) and the rates in MK2206 group (54% ±6%), FOXO3a-siRNA group (18%±7%) and Bim-siRNA group (11%±6%) were also higher than the control group.Compared with the triptolide group, the inhibition rate in MK2206 group was significantly increased, but decreased in FOXO3a-siRNA group and Bim-siRNA group(P<0.05). Conclusion: Triptolide induces T24 cells apoptosis through FOXO3a-Bim signaling pathway.

Metformin's effect on 5-fluorouracil,cisplatin,paclitaxel in laryngocarcinoma Hep-2 cells

Objective:To investigate metformin's effect on chemosensitivity of chemotherapeutic drug 5-fluorouracil in laryngocarcinoma Hep-2 cells. Investigate the variation trend of protein expression of AMPK pathway in the combined effect.Method:Laryngocarcinoma Hep-2 cells were treated with different concentrations of 5-fluorouracil in vitro together with or without metformin for 72 h. Use MTT assay to investigate the influence on the inhibition rate to Hep-2 cells. Hep-2 cells were treated with cisplatin, 5-fluorouracil or paclitaxel with or without metformin. Use Western blot assay to investigate the expression level of AMPKα, P21 or Cyclin D1 protein. Result:5-fluorouracil and metformin could inhibit the proliferation of Hep-2 cells. 5-fluorouracil in low concentration combined with metformin could increase the proliferation inhibition rate of Hep-2 cells. In the circumstances of using 5-fluorouracil in high concentration with metformin , the cell proliferation inhibition rate of combining group makes no differences with the single-drug group. The combination of metformin and 5-fluorouracil produced an antagonism action in Hep-2 cells.Western blot assay showed that metformin, cisplatin, 5-fluorouracil could have caused the increase of expression level of AMPK-α, P21 and Cyclin D1 in Hep-2 cells while Paclitaxel could have cause the decrease of expression level of Cyclin D1. Using combined drug could cause the change of protein expression. Conclusion:5-fluorouracil has been found to inhibit the proliferation of Hep-2 cells. Metformin has an antagonism on the anticancer effect to 5-fluorouracil in Hep-2 cells, and this antagonistic effect occurred partially through molecular signal pathways of AMPK-α, P21 and Cyclin D1 and it's significantly related to the cell cycle arrest.

Synthesis and the interaction of 2-(1H-pyrazol-4-yl)-1H-imidazo[4,5-f][1,10]phenanthrolines with telomeric DNA as lung cancer inhibitors

A novel series of 2-(1H-pyrazol-4-yl)-1H-imidazo[4,5-f][1,10]phenanthrolines were designed, synthesized and evaluated for their antitumor activity against lung adenocarcinoma by CCK-8 assay, electrophoretic mobility shift assay (EMSA), UV-melting study, wound healing assay and docking study. These compounds showed good inhibitory activities against lung adenocarcinoma. Especially compound 12c exhibited potential antiproliferative activity against A549 cell line with the half maximal inhibitory concentration (IC50) value of 1.48 μM, which was a more potent inhibitor than cisplatin (IC50 = 12.08 μM) and leading compound 2 (IC50 = 1.69 μM), and the maximum cell inhibitory rate being up to 98.40%. Moreover, further experiments demonstrated that compounds 12a–d can strongly interact with telomeric DNA to stabilize G-quadruplex DNA with increased ΔTm values from 12.44 to 20.54 °C at a ratio of DNA to compound 1:10. These results implied that growth inhibition of A549 cells mediated by these phenanthroline derivatives is possibly positively correlated to the fact their interaction with telomeric G-quadruplexs.

吴茱萸碱抑制Wnt/β-catenin信号通路诱导骨肉瘤MG-63细胞凋亡

Objective To investigate the effects and mechanism of evodiamine on the proliferation and apoptosis of osteosarcoma MG-63 cells. Methods MG-63 cells were cultured with evodiamine for 24 hours, and the cell proliferation was evaluated by methyl thiazolyl tetrazolium (MTT) assay. Cell cycle arrest, apoptosis and intracellular Ca2+ accumulation were evaluated by flow cytometry. BALB/C mice model of osteosarcoma was established to investigate the tumor inhibitory effect of evodiamine on human osteosarcoma. Wnt/β-catenin signaling protein expressions in osteosarcoma cells were detected by Western blotting. Results As concentration of evodiamine increasing (0.25 μmol/L, 0.5 μmol/L, 1.0 μmol/L, 2.0 μmol/L and 4.0 μmol/L), the inhibition rate of MG-63 cells increased [(4.18±1.26)%, (15.49±2.26)%, (40.55±6.57)%, (49.87±7.69)% and (60.42±8.29)%]. The difference was statistically significant between 2.0 μmol/L group and the control group (t=-2.66, P<0.05). MG-63 cells were cultured with 2.0 μmol/L evodiamine for 24 hours, and the apoptotic rate was (64.67±8.63)%, the proportion of S phase cells was (85.33±9.31)%, the fluorescence of Ca2+ was 97.33±21.31. The corresponding data of the control group were (4.94±0.81)%, (43.67±8.92)% and 28.67±8.92, the differences were statistically significant (t=-11.90, P<0.05; t=-7.22, P<0.05; t=-6.65, P<0.05). On mice model, the tumor weight of evodiamine group and the control group was (2.15±0.35)g and (4.29±0.49)g respectively, the difference was statistically significant (t=7.95, P<0.05). Comparing with the control group (1.00±0.00), evodiamine decreased the expression of β-catenin protein (0.72±0.36) and increased the expressions of Bax (1.15±0.27) and Caspase-3 (1.46±0.18) protein, and the differences were statistically significant (t=-3.05, P<0.05; t=-6.42, P<0.05; t=-5.85, P<0.05). Conclusion Evodiamine inhibits proliferation and induces apoptosis of human osteosarcoma MG-63 cells by blocking Wnt/β-catenin signaling. Key words: Evodiamine; Osteosarcoma; Cell proliferation; Apoptosis; Wnt/β-catenin