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Msi1基因沉默对人肝癌HepG2细胞的影响 被引量:4
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作者 王燕 章建国 +2 位作者 张曙 靳钦 卞婷婷 《交通医学》 2014年第2期115-118,共4页
目的:探讨siRNA沉默Msi1基因表达后,人肝癌HepG2细胞的增殖与凋亡的情况。方法:设计合成针对Msi1 mRNA序列的siRNA多条序列,分别转染人肝癌HepG2细胞。使用RT–PCR方法检测各组细胞Msi1基因表达情况;Western blot法检测survivin蛋白、ca... 目的:探讨siRNA沉默Msi1基因表达后,人肝癌HepG2细胞的增殖与凋亡的情况。方法:设计合成针对Msi1 mRNA序列的siRNA多条序列,分别转染人肝癌HepG2细胞。使用RT–PCR方法检测各组细胞Msi1基因表达情况;Western blot法检测survivin蛋白、caspase3蛋白表达情况;MTT法、平皿克隆实验检测HepG2细胞增殖情况;流式细胞术检测各组HepG2细胞凋亡情况。结果:(1)RT-PCR结果:转染后24 h后,序列a,b,c转染效率分别为65%,64%及62%。其抑制率分别是Msi1 siRNAa(68%)、Msi1 siRNAb(56%)、Msi1 siRNAc(35%)。(2)流式细胞术检测各组细胞凋亡:空白组、阴性组、脂质体组、Msi1 siRNAa组凋亡率分别为(4.14±0.26)%、(4.51±0.78)%、(4.19±1.21)%,(16.8±0.26)%,Msi1 siRNAa组与空白对照组比较差异有统计学意义(P<0.05)。(3)Msi1 siRNAa最能有效抑制肝癌HepG2细胞中Msi1表达,与空白对照组比较,Msi1 siRNAa组HepG2细胞生长、增殖速度明显减缓(P<0.05);survivin蛋白表达水平显著下调(P<0.05),而caspase3蛋白表达水平上调(P<0.05);Msi1 siRNAa组凋亡率增高(P<0.05)。结论:沉默Msi1可抑制人肝癌HepG2细胞的增殖,诱导其凋亡。 展开更多
关键词 原发性肝癌 Msi1基因 凋亡 增殖 HepG2细胞 逆转录聚合酶链反应 Western BLOT检测 流式细胞术 Msi1 reverse transcription-polymerase Chain Reaction (rt-pcr) Flow Cytometry(FCM)
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Abnormal expression of VEGF and its gene transcription status as diagnostic indicators in patients with non-small cell lung cancer 被引量:1
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作者 Yun Shi Yang Shi +4 位作者 Xuli Yang Jianrong Chen Qi Qian Dengfu Yao Guangzhou Wu 《Oncology and Translational Medicine》 CAS 2015年第5期201-207,共7页
Objective Angiogenesis is known to be essential for the survival,growth,invasion,and metastasis of lung cancer cells. Vascular endothelial growth factor(VEGF) is an important factor regulating angiogenesis of non-smal... Objective Angiogenesis is known to be essential for the survival,growth,invasion,and metastasis of lung cancer cells. Vascular endothelial growth factor(VEGF) is an important factor regulating angiogenesis of non-small cell lung cancer(NSCLC); however,its pathologic features and significance are unclear. In this study,the tissue VEGF expression levels and its gene transcriptional status,as well as circulating VEGF levels,were investigated in patients with lung disease. Methods VEGF protein and m RNA expression levels in 38 lung tissue samples were analyzed by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR),respectively. Circulating VEGF levels were detected quantitatively by an enzyme linked immuno-sorbent assay. Results The level of VEGF expression was significantly higher in lung cancer tissue than in the corresponding paracancerous or non-cancerous tissues. The average level of VEGF-positive staining was 76% in tissue samples from NSCLC patients; the levels were 89% in tissue samples from stage III patients and 92% in stage IV patients. High VEGF expression was also evident in cases with lymph node metastasis(84%),distant metastasis(90%),and lower differentiation degree(89%). VEGF m RNA in cancerous tissues was represented predominantly by the VEGF121 and VEGF165 isoforms. Circulating VEGF levels were significantly higher in NSCLC patients [(840 ± 324) pg/m L] than in patients with benign lung diseases [(308 ± 96) pg/m L] or in healthy individuals serving as controls [(252 ± 108) pg/m L]. Conclusion The over-expression of lung VEGF and its gene transcription status should be useful molecular indicators for NSCLC diagnosis. 展开更多
关键词 lung cancer vascular endothelial growth factor (VEGF) VEGF mRNA reverse transcription-polymerase chain reaction (rt-pcr IMMUNOHISTOCHEMISTRY
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Expression of CK19 mRNA and MUC-1 mRNA in the peripheral blood of patients with colorectal cancer and their clinical significances
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作者 Qian Wang Jie Liu Zewu An 《The Chinese-German Journal of Clinical Oncology》 CAS 2014年第7期316-319,共4页
Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to dis... Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to discuss the feasibility of colorectal carcinoma micro-metastasis detection of molecular markers. Methods: The expression level was detected by nested RT-PCR in 20 healthy people, 20 patients with colorectal adenoma and 90 cases of patients with colorectal cancer disease peripheral blood CK19 mRNA and MUC-1 mRNA. Results: The positive expression rate of CK19 mRNA and MUC-1 mRNA were: 58.89%(53/90) and 52.22%(47/90). No CK19 mRNA healthy people 20 cases in the control group in the peripheral blood, the expression of MUC-1 mRNA in 12 cases, the expression rate of 60%(12/20). In 20 cases of colorectal adenoma diseases have the expression of CK19 mRNA in 1 cases, the expression rate of 5%(1/20), the expression of MUC-1 mRNA in 10 cases, the expression rate of 50%. Patients with colorectal cancer CK19 mRNA, MUC-1 mRNA expression rate was significantly correlated with tumor staging, the degree of differentiation of the tumor cells and tumor metastasis(P < 0.05). Conclusion: Marker CK19 mRNA as the detection of micro-metastasis in peripheral blood of patients with colorectal cancer has good sensitivity and specificity, but CK19 mRNA, MUC-1 mRNA can be used to judge the prognosis of patients with colorectal cancer index. 展开更多
关键词 colorectal cancer (CRC) peripheral blood CK19 mRNA MUC-1 mRNA nested reverse transcription-polymerase chainreaction (Nested rt-pcr)
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Molecular characterization of yolk proteins in the female crab Neptunus pelagicus(A.Milne-Edwards,1861) from the Mediterranean Sea of Alexandria,Egypt
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作者 IBRAHIM G ELGHAZALY M 《Journal of Oceanology and Limnology》 SCIE CAS CSCD 2020年第2期438-453,共16页
This study aimed to characterize the morphological changes in the ovary of the female crab Neptunus pelagicus and to identify specific fractions of vitelloginin and vitelline molecules during primary and secondary vit... This study aimed to characterize the morphological changes in the ovary of the female crab Neptunus pelagicus and to identify specific fractions of vitelloginin and vitelline molecules during primary and secondary vitellogenesis.Samples of the blue crab were collected from the Mediterranean Sea of Alexandria monthly during 2017.Ovaries and oocytes in primary and secondary vitellogenesis were detached and treated for histological test.Native polyacrylamide gel electrophoresis(PAGE)Bis-Tris Gels was applied to identify vitelloginin(VN)and vitelline(VL)molecules.Protein Analyses were done by PAGE-SDS.The initial degenerate primers were built regarding the conserved amino acid domains of the yolk proteins.Primary and secondary vitellogeneses consisted of 8 phases.Lipoprotein fraction with molecular weight 550 kDa was identified in the hemolymph in secondary vitellogenesis.Two protein fractions(VLI&VLII)were identified in secondary vitellogenic oocytes.The electrophoresis performed with extract of stage I oocyte showed two protein fractions with molecular weights 550 kDa and 460 kDa.In stage II and III oocyte,4 subunits were presented of 180,195,140 and 120 kDa in VLI and 2 subunits with molecular weight of 110 kDa and 95 kDa in VLII.Another two fractions in stage V oocyte presented with molecular weights of 380 kDa and 360 kDa.Western blot analysis proved that both fractions were of four major polypeptide subunits with molecular weight of 180,125,90 and 85 kDa in each of the two VLs.The hybridization signal obtained by the Northern blot was detected in the hepatopancreas during ovarian cycle and in the ovary during secondary vitellogenesis.The result of the reverse transcription-polymerase chain reaction(RT-PCR)analysis showed that the mRNA that encodes the C-terminal region of the VN cDNA was found in the ovary in secondary vitellogenesis and in the hepatopancreas. 展开更多
关键词 Neptunus pelagicus VITELLOGENESIS vitelloginin and vitelline native polyacrylamide gel electrophoresis(PAGE)Bis-Tris Gels PAGE-SDS western blot analysis reverse transcription-polymerase chain reaction(rt-pcr)
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Predictive biomarker and clinicopathological characteristics analysis for esophageal cancer
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作者 Guogui Sun Wanning Hu +1 位作者 Jun Zhang Congrong Yang 《The Chinese-German Journal of Clinical Oncology》 CAS 2012年第2期83-88,共6页
Objective:The aim of this study was to explore the expression of manganese superoxide dismutase(MnSOD) in esophageal squamous cell carcinoma and its relationship with clinicopathological characteristics and its biolog... Objective:The aim of this study was to explore the expression of manganese superoxide dismutase(MnSOD) in esophageal squamous cell carcinoma and its relationship with clinicopathological characteristics and its biological behavior.Methods:Immunohistochemical method(SP method),reverse transcription-polymerase chain reaction(RT-PCR) and Western blot were combined to detect the MnSOD protein and mRNA expression in 45 cases of esophageal squamous cell carcinoma and the normal tissue that was 5 cm apart from the edge of esophageal cancer lesion and without documented microscopic invasive cancer.Meanwhile,analysis was performed on the relationship between the pathological features of esophageal cancer and its biological behavior.Results:In esophageal squamous cell carcinoma and normal esophageal tissue,MnSOD protein expression was identified in 31.1%(14/45) and 86.7%(31/45)(P = 0.003),respectively,with the relative expression levels of MnSOD mRNA were 0.310 ± 0.036 and 0.482 ± 0.053(P = 0.000).The longer the lesions and the deeper the invasion,the differentiation would become poorer and the expression level of MnSOD would get lower,indicating that the level of MnSOD protein and mRNA expression were closely related to esophageal squamous cell carcinoma in the length of lesion,depth of invasion,and degree of differentiation(P < 0.05).Nevertheless,it showed no association with the presence of the lymph node metastasis,lesion site and the macroscopic classification(P > 0.05).Conclusion:The MnSOD protein and mRNA expression were both decreased in esophageal squamous cell carcinoma tissue.This may be related to the carcinogenesis and development of esophageal cancer.Detection of the expression of MnSOD would be of clinical significance in understanding the prognosis and guiding therapeutic strategy of esophageal cancer. 展开更多
关键词 manganese superoxide dismutase(MnSOD) reverse transcription-polymerase chain reaction(rt-pcr esophageal squamous cell carcinoma IMMUNOHISTOCHEMISTRY
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Effects of Mangiferin on the Expression of TNF-α,iNos,ICAM-1 and Its mRNA in the Heart,Brain and Kidneys of SHR
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作者 Xiaoqin HU Xuewen ZENG +3 位作者 Erwei HAO Zhengcai DU Bei ZHOU Jiagang DENG 《Medicinal Plant》 CAS 2018年第2期35-39,77,共6页
[Objectives] To study the effects of Mangiferin( MGF) on TNF-α,iNOS,ICAM-1 and its mRNA expression in the heart,brain and kidneys of spontaneously hypertensive rats( SHR),and reveal the mechanism of its anti-inflamma... [Objectives] To study the effects of Mangiferin( MGF) on TNF-α,iNOS,ICAM-1 and its mRNA expression in the heart,brain and kidneys of spontaneously hypertensive rats( SHR),and reveal the mechanism of its anti-inflammatory injury in hypertension target organs.[Methods]SHRs were randomly divided into 5 groups: the model group,the high-dose,medium-dose,low-dose MGF groups and the Benazepril group,with 8 rats in each group,WKY rats were used for the normal control group. Besides,on-invasive blood pressure( BP) instruments were used to measure systolic blood pressure in the rats' tail artery,western blot was used to analyze the expression of TNF-α,iNOS,ICAM-1 and reverse transcription-polymerase chain reaction( RT-PCR) was used to analyze the expression of TNF-α,iNOS,ICAM-1 mRNA.[Results]Compared with the normal control group,the model group's BP level was significantly increased( P <0. 01)),but the MGF had no significant lowering BP effect( P > 0. 05); compared with the normal control group,the expression of TNF-α,iNOS,ICAM-1 and its' mRNA in the model group was significantly increased( P < 0. 05 or P < 0. 01),and MGF could reduce the level of expression of these inflammatory cytokines( P < 0. 05 or P < 0. 01); between the Benazepril group and high-dose,medium-dose,low-dose MGF groups,most of the indicators had no significant difference( P > 0. 05). [Conclusions]MGF had no significant lowering BP effect,SHR showed inflammatory injury in the heart,brain and kidneys,MGF showed improvement on the inflammatory injury,and the anti-inflammation mechanism may be associated with lowering TNF-α,i NOS and ICAM-1 and its mRNA expression. 展开更多
关键词 MANGIFERIN Spontaneously hypertensive rats(SHRs) Target organ Inflammatory factor reverse transcription-polymerase chain reaction(rt-pcr) Western blot
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