ABSTRACT
Intrahepatic cholangiocarcinoma (ICC) is a primary hepatic malignancy that originates from epithelial cells of bile duct. Lack of diagnostic measures and therapies leads to an increasing number of deaths from ICC worldwide. Here we described a case of 61-year-old Chinese female, who initially presented with right upper quadrant pain, combined with the results that a low density mass accompanied by multiple nodules occupied the right liver lobe by CT-scan, which also showed an aberrant right hepatic artery that participated in the right liver lobe and origining from the superior mesenteric artery, this patient was clinically considered as hepatic abscess (HA). The patient’s right upper quadrant pain was alleviated after been treated with the infusion chemotherapy of the aberrant right hepatic artery (ARHA) via percutaneous femoral arterial catheterization by Seldinger technique (Meropenem 7 days) following the failure of the liver-puncture drainage. However, the right upper quadrant pain occurred again 6 days later, serum CA19-9>1000.0 U/ml, which indicated the possibility of hepatic malignancy, so we performed laparotomy. The histopathological result of intraoperative frozen section demonstrated cholangiocarcinoma, unfortunately, it was unresectable. Finally, right lower lung pneumonia and pleural empyema happened to her and she succumbed to respiratory failure 22 days following surgery. In this report, we will discuss the case with reference to the literature.
ABSTRACT
Atherosclerosis (AS) is a common vascular disease, which can cause apoptosis of vascular endothelial cells. Notoginsenoside R1 (NGR1) is considered an anti-AS drug. MicroRNAs (miRNAs) are believed to play a vital role in cell apoptosis and angiogenesis. This study aimed to explore the mechanism of NGR1 for treating AS through miRNAs. Flow cytometry was used to detect the apoptosis rate. The levels of inflammatory cytokines interleukin (IL)-6 and IL-1β were detected using ELISA. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels were measured using corresponding assay kits. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed to detect miR-221-3p expression. Dual-luciferase reporter and RNA immunoprecipitation assays were carried out to examine the relationship between miR-221-3p and toll-like receptors 4 (TLR4). Also, western blot analysis was performed to determine the levels of TLR4 and nuclear factor kappa B (NF-κB) signaling pathway-related proteins. Oxidized low-density lipoprotein (ox-LDL) induced human umbilical vein endothelial cells (HUVECs) apoptosis, inflammation, and oxidative stress. NGR1 alleviated the negative effect of ox-LDL through promoting the expression of miR-221-3p in HUVECs. TLR4 was a target of miR-221-3p, and its overexpression could reverse the inhibition effects of miR-221-3p on apoptosis, inflammation, and oxidative stress. NGR1 improved miR-221-3p expression to inhibit the activation of the TLR4/NF-κB pathway in ox-LDL-treated HUVECs. NGR1 decreased ox-LDL-induced HUVECs apoptosis, inflammation, and oxidative stress through increasing miR-221-3p expression, thereby inhibiting the activation of the TLR4/NF-κB pathway. This study of the mechanism of NGR1 provided a more theoretical basis for the treatment of AS.
Subject(s)
Humans , Apoptosis/drug effects , Oxidative Stress/drug effects , Ginsenosides/pharmacology , MicroRNAs/adverse effects , Human Umbilical Vein Endothelial Cells/drug effects , Inflammation/metabolism , Lipoproteins, LDL/metabolism , Enzyme-Linked Immunosorbent Assay , Signal Transduction , Transcriptional Activation , Up-Regulation , Blotting, Western , NF-kappa B/antagonists & inhibitors , Reactive Oxygen Species , MicroRNAs/metabolism , Immunoprecipitation , Toll-Like Receptor 4/antagonists & inhibitors , Human Umbilical Vein Endothelial Cells/metabolism , Real-Time Polymerase Chain ReactionABSTRACT
DNA damage response (DDR) in different cell cycle starus of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated.The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA).The apoptotic ratio and the phosphorylation H2AX (S139)were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray.The expressions of γH2AX,Bcl-2,caspase-3 and caspase-9 were detected by Western blotting.DDR in 293T cells was detected after H2AX was silenced by RNAi method.Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment.The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P＜0.05).The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased.No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls.It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates.γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.
ABSTRACT
This study examined the role of regulated upon activation normal T cell expressed and secreted (RANTES) and its receptor C-C chemokine receptor type 5 (CCR5) in gastric cancer metastasis and the associated mechanism.The expression of RANTES and CCR5 was detected by using immunohistochemical staining and Western blotting in the gastric cancer tissues obtained from 60 gastric cancer patients with or without lymph node metastasis (n=30 in each).The results showed that the expression levels of RANTES and CCR5 were higher in gastric cancer with lymph node metastasis than in that without metastasis (P＜0.05).The expression levels of RANTES in 30 lymph nodes with cancerous invasion were higher than in 30 normal lymph nodes (P＜0.05).Chemotactic test revealed that the number of migrating gastric cancer cells (n=295.0±54.6) induced by the protein of cancer-invading lymph nodes was greater than that by the protein mixture from cancer-invading lymph nodes and RANTES antibody (n=42.5+11.6) (P＜0.05).RT-PCR showed that the expression levels of the main Th1 cytokines (IL-2,γ-IFN) were lower in gastric cancer with lymph node metastasis (2.22±0.90,3.26±1.15 respectively)than in that without metastasis (3.07±1.67,4.77±1.52 respectively) (P＜0.05),but the expression level of the main Th 2 cytokine (IL-10) was higher in gastric cancer with lymph nodes metastasis (6.06±2.04)than in that without metastasis (4.88±1.87) (P＜0.05).It was concluded that RANTES and its receptor CCR5 may contribute to gastric cancer metastasis through influencing the balance of Th1/Th2.RANTES and CCR5 may become a marker of gastric cancer metastasis.
ABSTRACT
Little is reported about the role of PTEN gene in the progression and prognosis of GISTs.This study examined the clinical implications of the tumor suppressor gene PTEN as a prognostic factor in the GISTs.Immunohistological staining and immunoblotting were employed to examine the PTEN protein expression,and its association with clinical measures.Clinicopathological features were reviewed by a retrospective examination of medical records.Reduced PTEN expression was significantly associated with tumor diameter,mitotic figure count,metastasis and pathological stage of tumor(P<0.05),and was not correlated with age,gender and tumor location(P>0.05).The 3-year survival rate of the patients with reduced PTEN expression was significantly lower than those with high PTEN expression(P<0.01).These results suggest that the expression of PTEN gene was significantly linked with the progression and metastasis of GISTs and it is an independent prognostic factor.
ABSTRACT
In order to examine the effect of GRIM 19 on colon cancer cell SW480, the recombinant adenovirus carrying GRIM19 gene was constructed and transfected into SW480 cells. GRIMI9 cDNA was amplified by PCR with the template pcxn2-GRlMl9 and cloned into the shuttle plasmid pAdTrack-CMV. The plasmid pAdTrack-CMV-GRIM19 was linearized by PmeI and homologously recombined with bone plasmid pAdEasy-1 in BJ5183, followed by identification by enzyme diges- tion. After transfection of linearized pAd-GRIM19 with PacI into HEK293 cells, Ad-GRIMI9 was obtained and amplified by 3 circles. SW480 cells were infected with Ad-GRIM19. The apoptosis rate was detected by flow cytometry. Agarose electrophoresis revealed the bands of recombinant plasmids identified by enzyme digestion were in the right range corresponding with expectation. Under the fluorescent microscopy, the package of Ad-GRIM19 in HEK293 cells and the expression of Ad-GRIM19 in SW480 cells were observed. The transfection of Ad-GRIM19 into SW480 cells in-creased the apoptosis rate of SW480 cells as compared with controls. It was concluded that Ad-GRIM19 was successfully constructed and the overexpression of GRIM19 in colon cancer cell lines could promote the apoptotic cell death.
ABSTRACT
In order to evaluate the effect of mitofusin-2 gene (mfn2) on proliferation and chemotherapy sensitivity of human breast carcinoma cell line MCF-7 in vitro, pEGFPmfn2 plasmid carrying full length of mitofusin-2 gene was transfected, by using sofast, into MCF-7 cells. Mitofusin-2 gene expression in MCF-7 cells transfected by sofast after 48 h was detected by PCR and Western blotting, and the stable expression of GFP protein in MCF-7 cells by Western blot analysis. The proliferation of MCF-7 cells was assayed by MTT and cell counting. By using PI method, the effects of mfn2 on the cell cycle distribution of MCF-7 were measured. Annexin-V/PI double labeling method was em- ployed to detect the changes in apoptosis induced by chemotherapeutics before and after transfection. The results showed that the MCF-7 cells transfected with mfn2 gene could stably and highly express GFP protein. MTT assay revealed that after transfection of mfn2 eDNA, the proliferation of MCF-7 cells was significantly inhibited. DNA histogram showed that cells arrested in S phase, and the per- centage of S phase cells was 42.7, 17.2 and 19.6 in mfn2 cDNA transfection group, blank plasmid transfection group and blank control group, respectively (P<0.05). The apoptosis ratio of the cells transfected with mfn2 gene was increased from 3.56% to 15.95%, that of the cells treated with camptothecin (CAMP) followed by mfn2 gene transfection was 69.6%, and that in blank plasmid transfection group and blank control group was 31.0% and 23.4% respectively (P<0.05). It was suggested that transfection of mfn2 gene could significantly inhibit the proliferation of MCF-7 cells and pro- mote their sensitivity to CAMP with a synergic effect.