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Objective:To investigate whether microRNA-30c (miR-30c) mediates the resistance of pancreatic cancer cells to gemcitabine (Gb) by targeting tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activating protein zeta polypeptide (YWHAZ) .Methods:SW1990 cell line with the lowest expression of miR-30c in human pancreatic cancer cell lines was screened by RT-qPCR. Gb-resistant cell line SW1990/Gb was established and divided into SW1990/Gb group (untransfected) , miR-30c over expression (Ad-miR-30c) group, Ad-miR-30c negative control (Ad-eGFP) group, and SW1990 group. The level of miR-30c was measured by RT-qPCR; the half inhibitory concentration (IC50) and drug resistance index (IR) were measured by CCK-8 method; the expression of drug resistance-related protein P-gp, apoptosis-related protein Caspase-1, migration and transfer-related proteins MMP-9, YWHAZ and downstream pathway-kinase mitogen-activated protein kinase (p38MAPK) /extracellular regulatory protein kinase 1 (ERK1) protein was measured by Western blot. After co-transfection of Ad-miR-30c and YWHAZ overexpressing adenovirus (Ad-YWHAZ) , the expression of P-gp and YWHAZ pathway related proteins was measured by Western blot method.Results:The IC50 (59.16±5.14, nmol/L) , IR (11.15±0.19) , expressions of YWHAZ protein (1.59±0.15) and P-gp (2.43±0.26) in SW1990/Gb-resistant cells were high, the expression of miR-30c (0.25 ±0.02) was low ( P<0.05) , and the p38MAPK/ERK pathway was activated. After up-regulating the expression of miR-30c (1.59±0.15) in SW1990/Gb cells, the IC50 (25.14±2.15, nmol/L) and IR (5.48±0.12) , YWHAZ (1.49±0.13) , P-gp (1.46± 0.10) decreased ( P<0.05) , and the p38MAPK/ERK pathway was activated. Up-regulating the expression of YWHAZ could reverse the above-mentioned effects of Ad-miR-30c ( P<0.05) . Conclusions:The expression of miR-30c is low in pancreatic cancer Gb-resistant cell lines. Up-regulating the expression of miR-30c can target and inhibit the YWHAZ/p38MAPK/ERK pathway, inhibit the expression of drug-resistant protein P-gp, and reduce the resistance of pancreatic cancer cells to Gb.
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Objective:To investigate the expression of immunoglobulin heavy constant gamma 1 (IGHG1) in pancreatic cancer and its effects on proliferation, migration and invasion of pancreatic cancer cell line PANC-1.Methods:From Jun. 2018 to Dec. 2020, 65 patients with pancreatic cancer who underwent surgical treatment at Xiang Yang No.1 People’s Hospital, Affiliated Hospital of Hubei University of Medicine were selected. The tumor tissue and normal adjacent tissues were removed, and the expression level of IGHG1 mRNA in pancreatic cancer tissues and adjacent tissues was detected by real-time quantitative PCR (RT-qPCR) . The correlation between IGHG1 expression and clinicopathological features of pancreatic cancer was analyzed. PANC-1 cells were divided into control group, IGHG1 negative control group (si-NC) and interference IGHG1 expression group (si-IGHG1) . IGHG1 mRNA expression, proliferation, migration, invasion and apoptosis of PANC-1 cells, as well as epithelial-mesenchymal transition (EMT) -related proteins E-cadherin and N-cadherin, Vimentin and Bax, Bcl-2, caspase3 protein expression were detected.Results:The expression level of IGHG1 mRNA in pancreatic cancer tissue was 2.47±0.23, significantly higher than 1.03±0.12 in adjacent tissues ( P<0.05) . the expression of IGHG1 was correlated with tumor differentiation, TNM stage and lymph node metastasis ( P<0.05) . Compared with the control group and si-NC group, the apoptosis rate of PANC-1 cells in the si-IGHG1 group was [ (5.34±0.65) %, (5.54±0.81) % vs (45.62±2.84) %]. Bax[0.34 ±0.05, 0.32±0.04 vs 1.13±0.12], caspase3 [0.43±0.05, 0.45±0.06 vs 1.22±0.13], and E-cadherin [0.78±0.12, 0.81±0.11 vs 1.34±0.08] protein expression levels increased significantly ( P<0.05) . The protein expression level was significantly increased ( P<0.05) , the expression level of IGHG1 mRNA [2.67±0.23, 2.61±0.21 vs 0.87±0.11] and protein [0.97±0.11, 1.01±0.10 vs 0.51±0.04], cell survival rate [ (98.21±0.56) %, (97.89±0.67) % vs (46.67±1.23) %], migration [ (76.12±2.72) %, (74.23±3.41) % vs (28.55±2.63) %] and invasion [ (85.32±3.71) %, 83.27±3.45) % vs (37.58±2.63) %] ability, and N-cadherin[1.12±0.13, 1.04±0.11 vs 0.61±0.08%], Vimentin[1.03±0.11, 0.97± 0.09 vs 0.49±0.07], and Bcl-2[0.87±0.08, 0.89±0.09 vs 0.62±0.07] protein expression levels were significantly reduced ( P<0.05) . Conclusions:The expression of IGHG1 is increased in pancreatic cancer, which is related to the occurrence and development of pancreatic cancer. Interference with IGHG1 expression can inhibit the proliferation, migration, invasion and EMT of pancreatic cancer cells.
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OBJECTIVE：To study the effects of gentiopicroside on the apo ptosis o f human pancreatic cancer cells PANC- 1，and to explore its mechanism from the perspective of IL- 6/JAK2/STAT3 signaling pathway. METHODS ：Using PANC- 1 cells as model ， the proliferation inhibition rate of cells was tested by MTT assay after treated with 0（negative contro ），2，4，8，16，32，64，128 mg/L gentiopicroside for 72 h and IC 50 were calculated. The cells were divided into negative control group ，gemcitabine group （positive control，4 mg/L）and gentiopicroside low-concentration ，medium-concentration and high-concentration groups （15，30，60 mg/L）. After cultured for 1，3，5，7 d，Trypan blue exclusion staining was used to count the survival cell ，and the growth of cells was investigated. After cultured for 72 h，colony formation assay was used to observe colony formation rate of cells ；the apoptotic rate of cells was detected by Hoechst 33258 staining；the mRNA and protein expressions of IL- 6，JAK2，STAT3 in cells were detected by RT-PCR and Western blotting assay. RESULTS ：4-28 mg/L gentiopicroside could inhibit the proliferation of cells （P＜0.05 or P＜ 0.01），in concentration dependent trend ；IC50 was 9.54 mg/L. Compared with negative control group ，survival cell count （cultured from 3，5，7 d），mRNA and protein expressions of IL- 6，JAK2 and STAT 3 in cells were decreased significantly in gemcitabine group ， gentiopicroside medium-concentration and high-concentration groups （P＜0.05 or P＜0.01），while the apoptotic rate was increased significantly （P＜0.01）. The colony formation rate of cellswere decreased significantly in gemcitabine group and gentiopicroside high-concentration group （P＜0.01）. mail：hb.gz@163.com Compared with gemcitabine group ，there was no statistical significance in above indexes of gentiopicroside high- 6716008。 concentration group （P＞0.05）. CONC LUSIONS：30，60 mg/L gentiopicroside could inhibit the proliferation and induce apoptosis of PANC- 1 cells，and 60 mg/L gentiopicroside is similar to gemcitabine in the effects. Its mechanism may be related to inhibiting the activation of IL- 6/JAK2/STAT3 signaling pathway.
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Objective To evaluate the regulation of the TRPV1 receptor on the expression of EGFR protein and the effects on related cell functions.Methods Human pancreatic cancer cells PANC-1 were cultured and treated by over-expression and siRNA.Cell proliferations were detected by CCK-8 experiment.RT-PCR test was used to detect the expression of onco-genes Akt2 and K-ras.Results EGFR protein was down-regulated by TRPV1 over-expression or by agonist activation in pan-creatic cancer cells.EGFR protein was increased by the interference of TRPV 1 expression.The proliferation rate of pancreatic cancer cells was decreased and Akt2,K-ras were significantly inhibited by TRPV1 over-expressing.Conclusion The expres-sion of TRPV1 in pancreatic cancer cells regulated the EGFR protein content.The cell proliferation and oncogene expression were inhibited by TRPV1 over-expressing.
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The present work is to study the chemical constituents from petroleum ether fraction of Tibetan medicine Swertia chirayita by column chromatography and recrystallization. The structures were identified by physical and chemical properties and spectral data as swerchirin (1), decussatin (2), 1,8-dihydroxy-3,5,7-trimethoxyxanthone (3), 1-hydroxy-3,5,7,8-tetramethoxyxanthone (4), bellidifolin (5), 1-hydroxy-3, 7-dimethoxyxanthone (6), methylswertianin (7), 1-hydroxy-3,5-dimethoxyxanthone (8), erythrodiol (9), oleanolic acid (10), gnetiolactone (11), scopoletin (12), sinapaldehyde (13), syringaldehyde (14), and β-sitosterol (15). Compounds 3, 4, 9, 11-14 were isolated from S. chirayita for the first time. Compounds 9 and 12 were firstly isolated from the genus Swertia. The cytotoxic activities of compounds 1, 2, 5, 7 and 8 against human pancreatic cancer cell lines SW1990 and BxPC-3,and the protective effects of these compounds against hydrogen peroxide (H2O2)-induced oxidative stress in human endothelium-derived EA.hy926 were investigated in vitro. The results showed no obvious effect at the high concentration of 50 μmol•L⁻¹.
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Objective To investigate the effects and mechanism of glucagon-like peptide-1 ( GLP-1 ) receptor agonist liraglutide on the proliferation and apoptosis of human pancreatic cancer cells. Methods The human pancreatic cancer cell line MIA PaCa-2 was incubated for 24 h with liraglutide at various concentrations (10-1 000 nmol/ L), or with 100 nmol/ L liraglutide for various durations (0-72 h). Cell proliferation was determined by Cell Counting Kit-8 (CCK-8) analysis. RT-PCR and Western blot were used to detect the mRNA and protein expression levels of related genes. Results GLP-1 receptor was expressed in the MIA PaCa-2 cells. Liraglutide suppressed cell proliferation, up-regulated the expression levels of pro-apoptotic protein Bax and down-regulated the expression levels of anti-apoptotic protein Bcl2 in human pancreatic cancer cells in a dose- and time-dependent manner. Meanwhile, liraglutide down-regulated the expression levels of insulin receptor (INSR) and insulin-like growth factor-1 receptor (IGF-1R), and the phosphorylation levels of their downstream signaling proteins Erk1 / 2 and Akt, in a dose- and time-dependent way. Conclusion Liraglutide inhibits proliferation and promotes apoptosis of human pancreatic cancer cells; the process may be mediated via suppressing the expression of INSR and IGF-1R and inhibiting activation of the downstream MEK/ Erk1 / 2 and PI3k/ Akt pathways.
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Objective: To observe the effect of monocrotaline (MCT) extract from Crotalaria sessiliflora on inducing human pancreatic cancer cell BxPC-3 into polyploid giant cells in vitro. Methods: BxPC-3 cells (1 × 104/mL) were inoculated with MCT (0, 5, 10, and 20 μg/mL) in RPMI-1640 for 72 h, respectively, then the cell morphology was observed by Giemsa staining, and the DNA content was measured by flow cytometry; BxPC-3 cells-induced apoptosis was checked by AnnexinV-FITC/PI double staining using flowcytometry, BxPC-3 polyploid giant cell genome was checked by chromosome spreading assay, and Cyclin B1 expression also was analysed by Western blotting. Results: Giemsa staining and DNA content by flow cytometry showed that MCT induced BxPC-3 cell into polyploid giant cells in vitro. MCT treatment for 72 h appeared 4N, and 8N polyploid giant of BxPC-3 cells, induction of apoptosis, decreased the expression of cyclin B1 in a dose dependent manner. Chromosome analysis demonstrated once again that polyploid giant cell was mainly in 8N. Conclusion: Monocrotaline might exert its antitumor effect by blocking the cell cycle, inducing apoptosis, and down-regulating Cyclin B1 protein.
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Objective: To investigate the influence of polysaccharide of Spirulina platensis(PSP) on inhibition activity of immobilized phycocyanin (PC) against proliferation of pancreatic cancer cells SW1990. Methods: PSP was added to the wells of cell culture polystyrene which was immobilized with PC, and SW1990 cells were cultured on this kind of biomaterial in serum free medium. Results: When the concentration of PSP was 15 mg/L, the inhibition activity was mainly caused by PSP, and when the concentrations were 10, 17.5 and 25 mg/L, PSP was cooperative with immobilized PC. The best immobilized amount of PC was 0.02 mg/well, which could inhibit 86% SW1990 cells. Conclusion: PSP can enhance the inhibition activity against pancreatic cancer cells SW1990 from immobilized PC.