Profilin 2 is highly expressed in gastric cancer and promotes tumor cell proliferation and migration / 南方医科大学学报

OBJECTIVE@#To investigate the expression of profilin 2 (PFN2) in gastric cancer and assess its potential value as a novel prognostic indicator and a therapeutic target.@*METHODS@#We collected gastric cancer and paired adjacent tissues from 100 patients for immunohistochemical detection of PFN2 expression. According to the expression level of PFN2, the patients were divided into two groups with high (46 cases) and low (48 cases) PNF2 expression in cancer tissues, and also into two groups with high (26 cases) and low (49 cases) PNF2 expression in adjacent tissues. Chi-square test, Spearman correlation and KaplanMeier survival analysis were used to analyze the relationship between PFN2 protein expression level and the patients' clinical parameters. We also tested the effects of PFN2 knockdown and overexpression on the proliferation and migration of MKN-45 cells using Transwell assay and CCK-8 assay.@*RESULTS@#The expression of PFN2 protein was significantly higher in gastric cancer tissues than in adjacent tissues (P < 0.01). PFN2 expression was positively correlated with M-stage of gastric cancer and VEGFR expression in the tumor tissues (P < 0.01). A high expression of PFN2 protein was significantly correlated with a poor prognosis of gastric cancer patients (P < 0.01), and was an independent predictor of the prognosis of gastric cancer. In MKN-45 cells, the cells overexpressing PFN2 showed significantly stronger proliferation and migration abilities than those with PFN2 knockdown (P < 0.001).@*CONCLUSION@#PFN2 protein is highly expressed in gastric cancer tissues to promote the proliferation and migration of the tumor cells. PFN2 may serve as a potential diagnostic marker, a prognostic indicator and a therapeutic target for gastric cancer.

Panax japlcus var polysaccharide regulates proliferation and apoptosis of gastric cancer MKN45 cells by targeting let-7a/CDK6 molecular axis / 中国肿瘤生物治疗杂志

@#Objective: To investigate the effect of panax japlcus var polysaccharide (PJPS) on the proliferation and apoptosis of gastric cancer MKN45 cells and its regulatory mechanism. Methods: Human gastric cancer cell lines (HGC27, MGC803, MKN45) and gastric mucosal epithelial cell line GES-1 were selected for this study. Let-7a mimics and let-7a inhibitor were transfected into MKN45 cells; Gastric cancer cell lines were treated with 100 μg/ml PJPS and MKN45 was selected as the subsequent experimental cell line. MKN45 cells were cultured with 0, 10, 50, 100 and 120 μg/ml PJPS, respectively. The proliferation and apoptosis rate of MKN45 cells were detected by CCK-8 and flow cytometry, respectively. Expressions of cell cycle dependent kinase 6 (CDK6) and apoptosis-related proteins in MKN45 cells were detected by Western blotting, and the expression level of miRNAs regulating the proliferation of gastric cancer cells was detectedbyReal-timequantitativePCR(qPCR).TheDualluciferasereportergeneassaywasusedtovalidatethetargeting relationship between let-7a and CDK6. Results: Compared with other gastric cancer cells, 100 μg/ml PJPS significantly inhibited the proliferation of MKN45 cells (P<0.01). At the same time, 100 μg/ml PJPS significantly up-regulated the expression of let-7a in MKN45 cells (P<0.01). The Dual luciferase reporter gene assay confirmed that CDK6 was the target gene of let-7a. Furthermore, PJPS inhibited the expression of CDK6 by up-regulating let-7a, thereby inhibiting the proliferation and inducing apoptosis of MKN45 cells (all P<0.01). Conclusion: PJPS inhibits proliferation and induces apoptosis of gastric cancer MKN45 cells by regulating the let-7a/ CDK6 axis.

Mechanism of Weichang' an on Autophagy of MKN45 Cells / 中国实验方剂学杂志

<b>Objective::To explore the effect of Weichang' an (WCA) on the autophagy of human gastric cancer MKN45 cells and its possible anti-cancer mechanism. <b>Method::MKN45 cells were cultured <italic>in vitro</italic> and incubated with different concentrations (250, 500, 1 000, 2 000 mg·L^-1) of WCA for 24, 48, 72 h. Cell counting kit-8 (CCK-8) assay was used to detect the cell proliferation. AO/EB Dyeing (AO) staining and monodansylcadaverin (MDC) staining were used to observe the changes of the effect of WCA on autophagosome and autophagic vesicles in gastric cancer MKN45 cells at 48 h. Real-time polymerase chain reaction (Real-time PCR) and Western blot were used to detect microtubule-associated protein 1 light chain 3 (LC3), Beclin1, sequestosome 1 (p62), human autophagy-related gene 5 (ATG5), human autophagy-related gene 7 (ATG7) mRNA and protein expression levels. <b>Result::WCA showed a dose-and-time-dependent growth inhibition at the concentration above 1 000 mg·L^-1. Compared with the blank group, WCA (500, 1 000, 2 000 mg·L^-1) significantly inhibited the proliferation of MKN45 cells (<italic>P</italic><0.05, <italic>P</italic><0.01). AO staining and MDC staining showed that autophagosomes and autophagic sacs increased with the rise of WCA concentration compared with the blank group. Real-time PCR and Western blot showed that the expressions of autophagy-related proteins LC3-Ⅱ, Beclin1, ATG5, ATG7, mRNA and protein increased gradually after WCA (1 000 mg·L^-1) intervention, while p62 expression decreased (<italic>P</italic><0.05, <italic>P</italic><0.01). <b>Conclusion::WCA induces the autophagy of human gastric cancer MKN45 cells in a time-and dose-dependent manner <italic>in vitro</italic>, which may be related to the up-regulation of LC3-Ⅱ, Beclin1, ATG5 and ATG7 as well as the down-regulation of p62 and LC3-Ⅰ.

The administration of peroxisome proliferator-activated receptors α/γ agonist TZD18 inhibits cell growth and induces apoptosis in human gastric cancer cell lines

Aim of Study: This study is to investigate the effects of a novel peroxisome proliferator-activated receptor (PPAR) α/γ dual agonist TZD18 on cell growth, apoptosis, caspase activity, mitochondrial membrane potential, cytochrome c release, and apoptotic-related protein expression in MKN-45 cells. Materials and Methods: 3-(4, 5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide assay against various human cancer cell lines was performed to investigate the whether TZD18 could in reduce the proliferation rates of cancer cells. The percentages of apoptotic cells and mitochondrial membrane potential level were determined by flow cytometry. The subcellular localization of cytochrome c was examined by immunofluorescence microscopy. Western blotting assay was performed to reveal the expression of apoptosis-related proteins. Results: The results showed that the administration of TZD18 could inhibit the growth of MKN-45 cells in a dose- and time-dependent manner. In addition, the apoptotic ratio increased sharply along with a significant increase of caspase activities, mitochondrial membrane potential, and cytochrome c release following TZD18 exposure. The expression of Bax and p27kip1 increased significantly, whereas the expression level of Bcl-2 protein was downregulated. Conclusion: These results indicated that the administration of PPAR α/γ agonist TZD18 may inhibit cell growth by inducing the apoptotic process in MKN-45 cells

Effects and mechanism of Rhizoma Paridis total saponin on invasion and migration abilities of human gastric adenocarcinoma cell MKN-45 induced by LiCl / 中草药

Objective: To investigate the effect and the possible mechanism of Rhizoma Paridis total saponin (RPTS) on human gastric cancer cell line MKN-45 proliferation, migration and invasion in vitro. Methods: MKN-45 cells were cultured in vitro and treated respectively with indicated concentrations of RPTS (2.5, 5.0, 10.0, 20.0, and 40.0 μg/mL) for 24 h, and cell viability of cell proliferation was detected by MTT assay; The invasive and metastatic ability of MKN-45 treated with indicated concentrations of RPTS (2.5, 5.0, 10.0 μg/mL) was detected by Transwell migration assay and wound healing assay; Elisa assay was employed to detect the concentrations of MMP-9 induced by LiCl after RPTS administration (10, 20, and 40 μg/mL) in the cell supernatant; Western blotting and qRT-PCR were respectively performed to investigate the invasion and migration related protein and mRNA level of VEGF, COX-2, and GSK-3β in RPTS-treated MKN-45 after LiCl stimulation for 24 h. Results: Compared with the control group, RPTS (10, 20, and 40 μg/mL) significantly inhibited the proliferation of MKN-45 cells (P < 0.05 and P < 0.001); RPTS (2.5, 5.0, 10.0 μg/mL) suppressed the invasion and migration of MKN-45 cells (P < 0.05 and P < 0.001); Compared with the model group, RPTS significantly downregulated the expression of MMP-9 in the cell supernatant of MKN-45 cells induced by LiCl (P < 0.05 and P < 0.01), and RPTS also decreased the protein and mRNA expression level of VEGF and COX-2, but it significantly upregulated the expression of GSK-3β at the protein and mRNA level (P < 0.05 and P < 0.001). Conclusion: RPTS play a pivotal role in suppressing the invasion and migration of MKN-45 cells in vitro, and its mechanism may be related to the regulating effects of the Wnt/β-catenin pathway in the human gastric adenocarcinoma cell.

Effect of Yangzheng Sanjie Decoction on Proliferation, Apoptosis and ERK Pathway of Human Gastric Cancer MKN-45 Cells / 中国实验方剂学杂志

Objective: To investigate the effect of Yangzheng Sanjie decoction on proliferation, apoptosis and extracellular signal-regulated kinase (ERK) pathway of human gastric cancer MKN-45 cells.Method: Gastric cancer cell line MKN-45 was treated for 24, 48, 72 h with Yangzheng Sanjie decoction (0.5, 1, 1.5, 2, 2.5, 3, 3.5 g·L-1); cell proliferation was measured by cell counting kit-8 (CCK-8); cell colony forming ability was observed by the plate cloning experiment after intervention with Yangzheng Sanjie decoction (0.4, 0.8 g·L-1); MKN-45 cells was treated with 4, 8 g·L-1, and then cell apoptosis was detected by flow cytometry; the expression of ERK and its phosphorylation level were detected by Western blot assay after treatment with 2, 4, 8 g·L-1.Result: Compared with the blank group, Yangzheng Sanjie decoction could significantly inhibit the proliferation of MKN-45 cells. After treatment for 24, 48 h, Yangzheng Sanjie decoction started from 2 g·L-1, and after treatment for 72 h, it started from 1.5 g·L-1, the cell viability gradually decreased in a concentration-dependent manner (PPP-1, cell colonies could not be formed; the apoptosis rate of Yangzheng Sanjie decoction was significantly higher than that of the blank group (PP-1, and the phosphorylation level of ERK protein in MKN-45 cells was down-regulated (PConclusion: Yangzheng Sanjie decoction can inhibit the proliferation of human gastric cancer cell line MKN-45 and promote its apoptosis. The mechanism may be related to the inhibition of phosphorylation of ERK.

Cytotoxic effect of double emulsion (w/o/w) cuso4 loaded pla nanoparticles on mkn-45 gastric adenocarcinoma cell line / Efecto citotóxico de nanopartículas de pla de doble emulsión (w/o/w) cargadas con cuso4 sobre la línea célular de adenocarcinoma gástrico mkn-45

Poly (L-lactic acid) (PLA) nanoparticles have the approval of the main institutions for drugs administration and therapeutics. However, the use of lactic acid polymer is controversial because lactic acid has been proposed as an energy source for cancer cells. The aim of this study was to evaluate the cytotoxic, apoptotic and cell cycle properties of PLA and CuSO4-loaded PLA biodegradable nanoparticles on MKN-45 gastric adenocarcinoma cell line. PLA nanoparticles for the delivery of the anticancer active principle CuSO4 were obtained using the double emulsion method. PLA and CuSO4 loaded PLA nanoparticles were morphologically characterized and their size determined using transmission electron microscopy (TEM). The cytotoxicity of this drug delivery system was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; apoptosis was evaluated using YO-PRO-1/Propidium Iodide and cell cycle analysis throughout flow cytometry. CuSO4-loaded PLA nanoparticles were effective inhibitors of MKN45 cancer cell growth. They increased cytotoxicity and apoptosis, and induced G1/Go cell cycle arrest;whereas the anticancer activity was increased using a 96 h treatment of a minimal (1mM) concentration of CuSO4 loaded in 40 µM PLA nanoparticles. The treatment with 40 µM lactic acid and PLA (40 µM) did not increase the rate of cell survival assays related to the control, which indicate that PLA use as a polymer carrier not induce proliferation of MKN-45 cancer cells. Our research presents novel data about the effect of PLA nanoparticles and CuSO4 on gastric cancer cell line MKN45.

Las nanopartículas de ácido poli L-láctico (PLA) tienen la aprobación de las principales instituciones de administración de medicamentos y terapéutica. Sin embargo, el uso de polímero de ácido láctico es controvertido ya que el ácido láctico se ha propuesto como una fuente de energía para las células cancerosas. El objetivo de este estudio fue evaluar las propiedades citotóxicas, la apoptosis y sobre el ciclo celular de las nanoparticulas de PLA biodegradable y de estas PLA nanopartículas cargadas con CuSO4 en la línea celular de adenocarcinoma gástrico MKN-45. Las nanopartículas de PLA para la administración del principio activo CuSO4 contra el cáncer se obtuvieron utilizando el método de doble emulsión. Las nanopartículas de PLA y PLA cargadas con CuSO4 se caracterizan morfológicamente y su tamaño fue determinaron usando microscopía electrónica de transmisión (TEM). Se evaluó la citotoxicidad de este sistema de administración de fármacos utilizando la 3 - (4,5-dimetiltiazol-2-il) -2,5-ensayo difeniltetrazolio (MTT); la apoptosis se evaluó usando yoduro de propidio/YO-PRO-1 y el análisis de ciclo celular por citometría de flujo. Las nanopartículas cargadas con CuSO4-PLA fueron eficaces inhibidores del crecimiento de las células MKN-45 cancerosas. Aumentaron citotoxicidad y la apoptosis, e inducen la detención del ciclo celular en G1/Go, mientras que la actividad contra el cáncer se incrementó con el uso de un tratamiento de 96 horas con una concentración mínima (1 mM) de CuSO4 cargado en nanopartículas con 40 µM de PLA. El tratamiento con 40 µM de ácido láctico y 40 µM PLA no aumentó la tasa de supervivencia de células en relación con el control, lo que indica que el uso de PLA como un polímero portador que no induce la proliferación de células de cáncer MKN-45. Nuestro estudio presenta nuevos datos sobre el efecto de las nanopartículas de PLA con CuSO4 en la línea celular de cáncer gástrico MKN-45.

Effects of CO_2 pneumoperitoneum on focal adhesion kinase of gastric cancer MKN-45 cells / 中华消化外科杂志

Objective To investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.