MiR-30e-5p overexpression promotes proliferation and migration of colorectal cancer cells by activating the CXCL12 axis via downregulating PTEN / 南方医科大学学报

OBJECTIVE@#To investigate the regulatory effects of miR-30e-5p on biological behaviors of colorectal cancer cells and the role of PTEN/CXCL12 axis in mediating these effects.@*METHODS@#Bioinformatic analysis was performed to explore the differential expression of miR-30e-5p between colorectal cancer tissues and normal tissues. RT-qPCR was used to detect the differential expression of miR-30e-5p in intestinal epithelial cells and colorectal cancer cells. Bioinformatics and dual luciferase assay were used to predict and validate the targeting relationship between miR-30e-5p and PTEN. Human and murine colorectal cancer cell lines were transfected with miR-30e-5p mimics, miR-30e-5p inhibitor, miR-30e-5p mimics+LV-PTEN, or miR-30e-5p inhibitor + si-PTEN. The changes in biological behaviors of the cells were detected using plate clone formation assay, CCK-8 assay, flow cytometry, scratch healing and Transwell assays. PTEN and CXCL12 expressions in the cancer cells were detected by Western blotting. The effects of miR-30e-5p inhibitor on colorectal carcinogenesis and development were observed in nude mice.@*RESULTS@#Bioinformatic analysis showed that miR-30e-5p expression was significantly elevated in colorectal cancer tissues compared with the adjacent tissue (P < 0.01). Higher miR-30e-5p expression was detected in colorectal cancer cell lines than in intestinal epithelial cells (P < 0.01). Dual luciferase assay confirmed the targeting relationship between miR-30e-5p and PTEN (P < 0.05). Transfection with miR-30e-5p mimics significantly enhanced proliferation and metastasis and inhibited apoptosis of the colorectal cancer cells (P < 0.05), and co-transfection with LV-PTEN obviously reversed these changes (P < 0.05). MiR-30e-5p mimics significantly inhibited PTEN expression and enhanced CXCL12 expression in the cancer cells (P < 0.01), and miR-30e-5p inhibitor produced the opposite effect. Transfection with miR-30e-5p inhibitor caused cell cycle arrest in the cancer cells, which was reversed by co-transfection with si-PTEN (P < 0.05). In the in vivo experiments, the colorectal cancer cells transfected with miR-30e-5p inhibitor showed significantly lowered tumorigenesis.@*CONCLUSION@#Overexpression of miR-30e-5p promotes the malignant behaviors of colorectal cancer cells by downregulating PTEN to activate the CXCL12 axis.

CXC chemokine receptor 4 regulates breast cancer cell cycle through S phase kinase associated protein 2 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of CXC chemokine receptor 4 (CXCR4) on cell cycle of breast cancer and its molecular mechanisms.</p><p><b>METHODS</b>The expression of CXCR4 and S phase kinase associated protein 2 (Skp2) was detected by real-time fluorescence quantitative PCR (fqRT-PCR) and Western blot in breast cancer cells. The expression of signal proteins and the downstream genes of Skp2 was detected by Western blot. The effect of CXCR4, PI3K/Akt pathway inhibitor LY294002 and ERK pathway inhibitor U0126 on cell cycle of breast cancer was detected by propidium iodide staining.</p><p><b>RESULTS</b>Skp2 was significantly down-regulated in CXCR4-downregulated cells and up-regulated in CXCR4-upregulated cells. CXCR4 also regulated the expression of Skp2 and other downstream genes by signaling protein. The proportion of cells in G/Gphase increased and that in S phase declined in CXCR4-downregulated cell, and the effect was more significant when combined with the use of LY294002 or U0126.</p><p><b>CONCLUSIONS</b>CXCR4 can affect cell cycle and inhibit the proliferation of breast cancer cells by regulating Skp2 gene expression through PI3K/Akt and ERK signaling pathway.</p>

Effects of lncRNA RP11-770J1.3 and TMEM25 expression on paclitaxel resistance in human breast cancer cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of long non-coding RNA(lncRNA) RP11-770J1.3 and transmembrane protein 25 (TMEM25) on paclitaxel resistance in human breast cancer MCF-7/PR cell line.</p><p><b>METHODS</b>The expression of lncRNA RP11-770J1.3 and TMEM25 in human breast cancer MCF-7(paclitaxel sensitive) and MCF-7/PR(paclitaxel resistant) cells were detected by quantitative RT-PCR. The synthetic interfering fragments of lncRNA RP11-770J1.3 and TMEM25 were transfected into MCF-7/PR cells. Sulforhodamine B assay was used to detect the sensitivity of MCF-7/PR cells to paclitaxel after interference of lncRNA RP11-770J1.3 and TMEM25. The expression of multidrug-resistance genes and proteins were detected by qRT-PCR and Western blot, respectively.</p><p><b>RESULTS</b>lncRNA RP11-770J1.3 and TMEM25 were highly expressed in MCF-7/PR cells, and were significantly down-regulated after transfection of synthetic interfering fragments. Down-regulation of lncRNA RP11-770J1.3 and TMEM25 enhanced the sensitivity of MCF-7/PR cells to paclitaxel, and inhibited the expression of MRP, BCRP and MDR1/P-gp (all<0.05). Such effects were more significant when lncRNA RP11-770J1.3 and TMEM25 were both down-regulated (all<0.05).</p><p><b>CONCLUSIONS</b>lncRNA RP11-770J1.3 and TMEM25 are highly expressed in MCF-7/PR cells, and the down-regulation of lncRNA RP11-770J1.3 and TMEM25 can enhance paclitaxel sensitivity in MCF-7/PR cells.</p>

Effects of lidocaine on peripheral blood mononuclear cells from patients with atopic dermatitis stimulated by the Staphylococcus aureus exotoxin TSST-1 / 中华皮肤科杂志

Objective To investigate the effect of lidocaine on Staphylococcus aureus exotoxin-stimulated peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis (AD).Methods Peripheral blood samples were collected from 6 patients with AD,and PBMCs were isolated by a routine method.Then,the PBMCs were stimulated by the Staphylococcus aureus exotoxin toxic shock syndrome toxin-1 (TSST-1) in the absence or presence of lidocaine at varying concentrations.The 3H-TdR incorporation method was performed to detect the proliferation of monocytes,and enzyme-linked immunosorbent assay (ELISA) to quantify the levels of T helper type 1 (Th1) and Th2 cytokines released by PBMCs.Human HaCaT keratinocytes were co-cultured with lidocaine-and TSST-1-stimulated PBMCs from patients with AD for 72 hours,then,Western blot was conducted to examine the expression of filaggrin protein in HaCaT cells.Results TSST-1 (100 μg/L) significantly enhanced the proliferation of PBMCs from patients with AD (stimulation index =75 ± 2.12,P ＜ 0.05),as well as the release of tumor necrosis factor-α (TNF-α),interferon (IFN)-γ,interleukin (IL)-2,IL-12,IL-4,IL-5 and IL-13 by the PBMCs (all P ＜ 0.05).Compared with the blank control group,100 μmol/L lidocaine significantly inhibited the TSST-1-stimulated proliferation of PBMCs from patients with AD (stimulation index =58 ± 3.14,P＜ 0.05),as well as the release of IL-4,IL-5,IL-13,TNF-α and IFN-γ by the stimulated PBMCs (all P ＜ 0.05).Western blot showed that 100 μmol/L lidocaine significantly blocked the down-regulation of filaggrin expression in HaCaT cells (P ＜ 0.01).Conclusion Lidocaine has a significant inhibitory effect on the activation of TSST-1-stimulated PBMCs from patients with AD.

Effect of berberine on proliferation and apoptosis of MCF-7 cells / 安徽医科大学学报

Objective To investigate the effect and mechanisms of the Berberine on the human breast adenocarci-noma cell line MCF-7 . Methods MTT method was used to evaluate the proliferation effect of MCF-7 and the per-centage of apoptotic cells were determined by flow cytometric analysis. The expressions of JAK2, p-JAK2, p-STAT3, STAT3, Bax, Bcl-2, Cleavage-PARP and Cleavage-Caspase3 were detected by Western blotting. Results The results showed that BBR treatment decreased the activation of JAK2/STAT3 signal pathway and up-regulated the expression of Bax, Caspase3 and PARP activation with the decrease of the expression of Bcl-2. Wherefore, ex-pression of the constitutively active form of STAT3 could attenuate the effect of BBR on the MCF-7 cell. Conclusion Berberine can induces apoptosis and proliferation inhibition of breast adenocarcinoma cell lines of MCF-7 through inhibition of the JAK2/STAT3 signaling pathway.

Effect of mitochondrial DNA deletion on growth and invasiveness of human lymphoma Namalwa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effect of mitochondrial DNA (mtDNA) deletion on the growth and invasiveness of human lymphoma Namalwa cells.</p><p><b>METHODS</b>ρ(0)-Namalwa cells with mtDNA deletion were generated by treating Namalwa cells with ethidium bromide and confirmed by selective ρ(0) test medium analysis, PCR and Western blotting. The growth of ρ(0)-Namalwa cells was evaluated by MTT assay and cell cycle analysis, and the cell migration and invasiveness were assessed with Transwell assay. Reactive oxygen species (ROS) production and cytosolic Ca(2+) were detected by flow cytometry.</p><p><b>RESULTS</b>ρ(0)-Namalwa cells could grow and divide normally in selective medium supplemented with uridine and pyruvate but not in nonselective medium. PCR did not yield the products of mtDNA, nor was COXII expression detected in ρ(0)-Namalwa cells. ρ(0)-Namalwa cells showed an obvious attenuation of cell proliferation and migration abilities with significantly lowered ROS production and cytosolic Ca(2+).</p><p><b>CONCLUSION</b>The suppressed growth and migration of ρ(0)-Namalwa cells may be the result of decreased ROS production and cytosolic Ca(2+).</p>

Effect of Oxaliplatin on cell cycle of hepatocellular carcinoma cell line HepG2 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of Oxaliplatin (L-OHP) on cell cycle in hepatocellular carcinoma cell line HepG2 and the involved mechanism.</p><p><b>METHODS</b>Inhibitory effect of L-OHP on the proliferation of HepG2 cells was determined by MTT assay. Cell cycle distribution was shown by flow FCM. The expression levels of cyclinD1, CDK2, CDK4, p16, p21, p53 were detected by RT-PCR and Western blot.</p><p><b>RESULTS</b>MTT method revealed that L-OHP inhibited proliferation of hepatocellular carcinoma HepG2 cells in a dose- and time-dependent manner. L-OHP induced S cell cycle arrest in HepG2 cell; down-regulated the levels of CDK4, cyclinD1 and up-regulated the levels of p21, p53. There were no significant changes of CDK2 and p16 after L-OHP treatment.</p><p><b>CONCLUSION</b>L-OHP inhibits the proliferation of HepG2 cells by blocking cell at S stage, which may be resulted from the activity of CDK4, CyclinD1 and p21.</p>

Suppression of breast cancer proliferation and induction of apoptosis via AKT and ERK1/2 signal transduction pathways by synthetic polypeptide derived from viral macrophage inflammatory protein II / 华中科技大学学报（医学）（英德文版）

SDF-1α, a ligand for the chemokine receptor CXCR4, is well known for mediating the migration of breast cancer cells. In a previous study we demonstrated that a synthetic 21-mer peptide antagonist of CXCR4 (NT21MP) derived from the viral macrophage inflammatory protein II could antagonize tumor growth in vivo by inhibiting cellular proliferation and inducing apoptosis in breast cancer cells. However, the role of SDF-1α in the signaling pathways underlying the proliferation of human breast cancer cells and associated signaling pathways and inhibiting signal pathways of NT21MP remained unclear. The present study investigated the mechanism of NT21MP on anti-tumor in breast cancer in vitro. The effect of NT21MP on the viability of cells was determined by the MTT assay. Annexin V-FITC and PI staining was performed to detect early stage apoptosis in SKBR3 cells treated with SDF-1α and AMD3100 or NT21MP. Western blotting techniques were used to assay the composition of phosphoproteomics and total proteins present in the SKBR3 breast cancer cells. RT-PCR and Western blotting technique were used to detect the effect of NT21MP and AMD3100 on Bcl-2 and Bax expression. The results indicated that SDF-1α prevented apoptosis and promoted the proliferation of SKBR3 human breast cancer cells. As compared with untreated SKBR3 cells, Treatment with SDF-1α significantly increased cell viability, and NT21MP abolished the protective effects of SDF-1α dose-dependently (P<0.05). There was a significant decrease in the percentage of apoptotic cells after SDF-1α treatment as compared with control group (2.7%±0.2% vs. 5.7%±0.4%, P<0.05). But pretreatment of SKBR3 cells with NT21MP significantly attenuated the antiapoptotic effects of SDF-1α as compared with SKBR3 cells without NT21MP pretreatment. The proliferative and anti-apoptotic effects of SDF-1α in SKBR3 cells were associated with an increase in AKT and ERK1/2 phosphorylation as well as a decrease in Bax expression and an increase in Bcl-2 expression. These changes in intracellular processes were blocked by NT21MP in a dose-dependent manner(P<0.05). In conclusion, NT21MP efficiently inhibits SDF-1α-induced proliferation and antiapoptosis in SKBR3 cells by reducing the levels of phosphorylated AKT and ERK1/2, as well as decreasing the ratio of expression of Bcl-2 relative to Bax.

Grape seed extract inhibit proliferation of breast cancer cell MCF-7 and decrease the gene expression of survivin / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of grape seed extract (GSE) on breast cancer cell MCF-7 about the of proliferation and the gene expression of survivin, and to investigate the molecular biological mechanism of inhibition by GSE.</p><p><b>METHOD</b>MTT was used to measure the role of proliferated inhibition of GSE at the dosage of 40, 80, 120, 160, 200 mg x L(-1) with different time. To calculate the IC50 of GSE and to select the suitable concentration and treatment time. The change of cell cycle was detected by flow cytometry. mRNA expression of Survivin was observed by RT-PCR. Luciferase kit was used to observe the change of core promoter of survivin which was inserted in the flourescence report vector. And further to analyse the bind side of transcription factor on the sequence of core promoter of survivin was further analysed by using the microsoft of bioinformatics.</p><p><b>RESULT</b>GSE could inhibit the proliferation of breast cancer cell MCF-7 with time and dosage-dependent (P < 0.05). GSE could arrest the cell cycle in S periods (P < 0.05) and also inhibit the mRNA expression of survivin effectively (P < 0.01); GSE could down-regulate the activity of core promoter of survivin significantly (P < 0.01).</p><p><b>CONCLUSION</b>GSE can inhibit the proliferation of breast cancer cell MCF-7 through arresting the cell cycle in S periods. The mechanism may be that GSE regulate the activity of transcription factor which are related to the activity of core promoter of survivin and decrease the gene expression of survivin.</p>

TrKA-siRNA inhibits the expression of NF-?B and promote apoptosis of breast cancer cell line MCF-7 / 基础医学与临床

Objective Investigate the effect of TrKA variation on the expression of neucleoprotein NF-?B P65 and apoptosis.Methods To construct the expression vector of TrKA small interfering RNA,the recombinant was transfected into MCF-7 cells.the stable cell line expressing TrKA small interfering RNA were selected by G418.The mRNA and protein of TrKA were tested by real-time PCR,Western-blot and Immunohistochemistry.The change of neucleoprotein NF-?B P65 was detected by WB,Flow cytometry was used to observe the cell apoptosis.ResultsThe expression vector of TrKA-siRNA was successfully constructed.The mRNA and protein of TrKA were decreased by 74.7% and 80.5% respectively(P

Construction of Survivin specific small interfering RNA expression vector,its antiproliferative and proapoptotic effect on lung adenocarcinoma cells / 基础医学与临床

Objective To test the antiproliferative and proapoptotic effect of Survivin small interfering RNA(siRNA) expression vector on lung adenocarcinoma cell A549 in vitro.Methods The Survivin-siRNA expression vector was constructed and conformed by sequencing.The inhibitory effect of Survivin-siRNA was tested by fluorescent quantitative reverse transcription polymerase chain reaction(FQ RT-PCR),Western blot and immunohistochemistry.The cell proliferation and apoptotic rate were assayed by tetrazolium bromide(MTT)colorimetry and flow cytometry.Results Survivin-siRNA expression vector was constructed and transfected into A549 cell.It effectively reduced mRNA and protein level of Survivin.Immunohistochemistry also showed lower expression of Survivin.The A549 cells transfected with Survivin-siRNA had a lower cellular growth rate than that of control group(P

The study of case-guide comprehensive experimente of clinical biochemistry / 中华医学教育探索杂志

Objective To reform experimental course teaching methods of clinical bio-chemistry and elevate students interesting and ability. Methods The reformed teaching methods was taken in clinical laboratory techniques and start case-guide comprehensive experiment of clinical biochemistry. Results The innovation of teaching methods of clinical biochemistry obtained satisfactory achievement,improved students thinking skills and overall quality. Conclusion It accorded with the trend of teaching innovation and was advantageous to increasing the comprehensive predisposition.

The experimental studies of the effects of As_2O_3 on the expression of telomerase hTERT-mRNA and apoptosis of HL-60 cells / 中国药理学通报

AIM To investigate the effects of apoptosis and the expression of telomerase hTERT mRNA activity of HL-60 cells treated with As 2O 3 in different concentrations. METHODS HL-60 cells was cultured with As 2O 3 in different concentrations for 48 hours, then, cell apoptosis was detected with flow cytometry and the expression of telomerase hTERT mRNA was determined with RT-PCR respectively. RESULTS The percentage of the apoptosis in HL-60 cells were 2 28%?0 40%?2 55%?0 22% and 47 57%?2 79% respectively, as the cells were treated with 0 1 ?mol?L -1,1 0 ?mol?L -1,5 0 ?mol?L -1 As 2O 3 for 48 hours. The difference was significant between 5 0 ?mol?L -1 and 0 1～1 0 ?mol?L -1 groups(P