Human trefoil factor 2 （hTFF2） is considered as one of the most important initiators of mucosal healing in the gastrointestinal tract by promoting cell migration and suppressing apoptosis. However, it is hard to obt...Human trefoil factor 2 （hTFF2） is considered as one of the most important initiators of mucosal healing in the gastrointestinal tract by promoting cell migration and suppressing apoptosis. However, it is hard to obtain hTFF2 from human tissue and many recombinant hTFF2 produced in vitro exist as fusion proteins. The purpose of the present study was to produce native hTFF2 while maintaining its biological activities. The open reading frame of hTFF2 was inserted into a pET-32a（＋） expression vector, and hTFF2-TRX fusion protein was successfully expressed in Escherichia coli and purified by Nickel-nitrilotriacetic acid affinity chromatography and reverse-phase HPLC steps. The recombinant fusion protein （purity〉95%） was cleaved by Factor Xa at 23 ~C to release hTFF2. After removal of Factor Xa and undigested fusion proteins, hTFF2 was purified and identified by SDS-PAGE and Western blotting. The yield of recombinant hTFF2 was about 5 mg/L. The recombinant hTFF2 could promote IEC-6 cells migration and in vitro wound healing via the activation of ERK1/2. Recombinant hTFF2 could also inhibit apoptosis of HCT-116 cells induced by 50 lamol/L ceramide In summary, our results showed that the recombinant hTFF2 was expressed in E. coli and successfully purified after cleavage with the fusion partner with high yield while maintaining its biological activities. Recombinant hTFF2 might be useful for investigating the molecular mechanism of hTFF2 and development of hTFF2-related drugs.展开更多
AIM: To investigate the expression of TFF2 and Helicobacter pyloriinfection in carcinogenesis of gastric mucosa.METHODS: The expression of TFF2 was immunohistochemically analyzed in paraffin-embedded samples from 119 ...AIM: To investigate the expression of TFF2 and Helicobacter pyloriinfection in carcinogenesis of gastric mucosa.METHODS: The expression of TFF2 was immunohistochemically analyzed in paraffin-embedded samples from 119 patients with endoscopic biopsy and subtotal gastrectomy specimens of gastric mucosal lesions, including 16 cases of chronic superficial gastritis (CSG), 20 chronic atrophic gastritis (CAG),35 intestinal metaplasia (IN), 23 gastric epithelial dysplasia (GED) and 25 gastric carcinoma (CA), and Helicobacter pylori infection was detected by Warthin-Starry staining.RESULTS: 1:TFF2 was located in the cytoplasm of gastrk mucous neck cell. The expression of TFF2 was 100 %,100 %, 0, 56.5 % and 0 in CSGs, CAGs, INs, GEDs and CAs, respectively. 2: The value of TFF2 positive cell density in CSG with Helicobacter pyloriinfection was higher than that without Helicobacter pyloriinfection. (52.89±7.27vs46.49±13.04, P＞0.05); But the value of TFF2 positive cell density in CAG and GED with Helicobacter pyloriinfection was significantly lower than that without Helicobacter pylori infection (18.17±4.09 vs 37.93±13.80, P＜0.01 and 14.44±9.32 vs 24.84±10.22, P＜0.05).CONCLUSION: Increase of TFF2 expression in CSG is perhaps associated with the protective mechanism after gastric mucosal injury. Decrease of TFF2 expression in CAG possibly attributes to the decrease in the number of gastric gland cell expressing TFF2. Re-expression of TFF2 in gastric epithelial dysplasia implies that TFF2 possibly contributes to the initiation of gastric carcinoma. The effect of Helicobacter pylori on the expression of TFF2 depends on the status of gastric mucosa.展开更多
目的探讨基质金属蛋白酶抑制基因(reversion-inducing cysteinerich protein with Kazal Motifs,RECK)、三叶因子2(Trefoil factors2,TFF2)、雌激素受体β亚型(ERβ)在子宫内膜癌中的表达及抑癌基因p16甲基化的临床意义。方法随机收集...目的探讨基质金属蛋白酶抑制基因(reversion-inducing cysteinerich protein with Kazal Motifs,RECK)、三叶因子2(Trefoil factors2,TFF2)、雌激素受体β亚型(ERβ)在子宫内膜癌中的表达及抑癌基因p16甲基化的临床意义。方法随机收集子宫内膜癌术后组织标本80例与同期术后正常子宫内膜组织标本45例,采用免疫组化法检测RECK、TFF2与ERβ的蛋白表达,MSP法检测p16甲基化。结果 RECK、TFF2、ERβ在子宫内膜癌组织中的阳性表达率分别为:50.00%、56.25%与48.75%,均显著低于正常组(82.22%、86.67%与80.00%),差异具有统计学意义(P<0.05);子宫内膜癌组TFF2、ERβ的阳性表达率均显著高于宫颈癌组(P<0.05),而子宫内膜癌组RECK、TFF2、ERβ的阳性表达率与术前化疗癌症组比较无显著性差异(均P>0.05)。子宫内膜癌组织中RECK、TFF2的阳性表达与临床分期、组织学分级、肌层浸润程度及淋巴结转移密切相关;ERβ的表达与组织学分级、肌层浸润程度及淋巴结转移密切相关,而与临床分期无相关性,子宫内膜癌组织存在p16基因全部或部分甲基化,正常子宫内膜组织不存在p16基因甲基化现象;其中子宫内膜癌组织中p16甲基化有42例(52.50%),p16甲基化与临床分期,组织学分级、肌层浸润与淋巴结转移密切相关。结论 RECK、TFF2、ERβ、p16甲基化在子宫内膜癌的不同组织学分级、肌层浸润程度与有无淋巴结转移中的表达各异,并在子宫内膜癌的诊治中具有临床应用价值。展开更多
To the Editor:Gastric cancer(GC)ranks third in incidence and mortality both worldwide and in China.[1,2]Intestinal metaplasia(IM)significantly increases risk of GC so that identifying high-risk IM patients who will pr...To the Editor:Gastric cancer(GC)ranks third in incidence and mortality both worldwide and in China.[1,2]Intestinal metaplasia(IM)significantly increases risk of GC so that identifying high-risk IM patients who will progress to GC is crucial.Currently,the effect of many risk-stratification methods for gastric precancerous lesions(GPLs)was minimal.Monoclonal gastric cancer 7 antigen(MG7-Ag)combined with cyclooxygenase-2 has been shown earlywarning value for the progression of GPLs.[3]The expression of human telomerase reverse transcriptase(hTERT)was proven to be related to state of cell proliferation in IM tissue.[4]Loss expression of trefoil factor family 2(TFF2)from spasmolytic polypeptideexpressing metaplasia to IM may lead to a hyperproliferation and deleterious mutations.[5]We tried to construct and verify a multimolecular prediction model included MG7-Ag and hTERT and TFF2,which could identify high-risk IM patients and have early-warning value for GC.展开更多
基金"973"项目(2010CB529800)国家基金委面上项目(81160302,30870304)+1 种基金中国科学院"西部之光"(Y102291081)the Science and Technology Department of Yunnan Province (2011C1139)~~
文摘Human trefoil factor 2 （hTFF2） is considered as one of the most important initiators of mucosal healing in the gastrointestinal tract by promoting cell migration and suppressing apoptosis. However, it is hard to obtain hTFF2 from human tissue and many recombinant hTFF2 produced in vitro exist as fusion proteins. The purpose of the present study was to produce native hTFF2 while maintaining its biological activities. The open reading frame of hTFF2 was inserted into a pET-32a（＋） expression vector, and hTFF2-TRX fusion protein was successfully expressed in Escherichia coli and purified by Nickel-nitrilotriacetic acid affinity chromatography and reverse-phase HPLC steps. The recombinant fusion protein （purity〉95%） was cleaved by Factor Xa at 23 ~C to release hTFF2. After removal of Factor Xa and undigested fusion proteins, hTFF2 was purified and identified by SDS-PAGE and Western blotting. The yield of recombinant hTFF2 was about 5 mg/L. The recombinant hTFF2 could promote IEC-6 cells migration and in vitro wound healing via the activation of ERK1/2. Recombinant hTFF2 could also inhibit apoptosis of HCT-116 cells induced by 50 lamol/L ceramide In summary, our results showed that the recombinant hTFF2 was expressed in E. coli and successfully purified after cleavage with the fusion partner with high yield while maintaining its biological activities. Recombinant hTFF2 might be useful for investigating the molecular mechanism of hTFF2 and development of hTFF2-related drugs.
文摘AIM: To investigate the expression of TFF2 and Helicobacter pyloriinfection in carcinogenesis of gastric mucosa.METHODS: The expression of TFF2 was immunohistochemically analyzed in paraffin-embedded samples from 119 patients with endoscopic biopsy and subtotal gastrectomy specimens of gastric mucosal lesions, including 16 cases of chronic superficial gastritis (CSG), 20 chronic atrophic gastritis (CAG),35 intestinal metaplasia (IN), 23 gastric epithelial dysplasia (GED) and 25 gastric carcinoma (CA), and Helicobacter pylori infection was detected by Warthin-Starry staining.RESULTS: 1:TFF2 was located in the cytoplasm of gastrk mucous neck cell. The expression of TFF2 was 100 %,100 %, 0, 56.5 % and 0 in CSGs, CAGs, INs, GEDs and CAs, respectively. 2: The value of TFF2 positive cell density in CSG with Helicobacter pyloriinfection was higher than that without Helicobacter pyloriinfection. (52.89±7.27vs46.49±13.04, P＞0.05); But the value of TFF2 positive cell density in CAG and GED with Helicobacter pyloriinfection was significantly lower than that without Helicobacter pylori infection (18.17±4.09 vs 37.93±13.80, P＜0.01 and 14.44±9.32 vs 24.84±10.22, P＜0.05).CONCLUSION: Increase of TFF2 expression in CSG is perhaps associated with the protective mechanism after gastric mucosal injury. Decrease of TFF2 expression in CAG possibly attributes to the decrease in the number of gastric gland cell expressing TFF2. Re-expression of TFF2 in gastric epithelial dysplasia implies that TFF2 possibly contributes to the initiation of gastric carcinoma. The effect of Helicobacter pylori on the expression of TFF2 depends on the status of gastric mucosa.
文摘目的探讨基质金属蛋白酶抑制基因(reversion-inducing cysteinerich protein with Kazal Motifs,RECK)、三叶因子2(Trefoil factors2,TFF2)、雌激素受体β亚型(ERβ)在子宫内膜癌中的表达及抑癌基因p16甲基化的临床意义。方法随机收集子宫内膜癌术后组织标本80例与同期术后正常子宫内膜组织标本45例,采用免疫组化法检测RECK、TFF2与ERβ的蛋白表达,MSP法检测p16甲基化。结果 RECK、TFF2、ERβ在子宫内膜癌组织中的阳性表达率分别为:50.00%、56.25%与48.75%,均显著低于正常组(82.22%、86.67%与80.00%),差异具有统计学意义(P<0.05);子宫内膜癌组TFF2、ERβ的阳性表达率均显著高于宫颈癌组(P<0.05),而子宫内膜癌组RECK、TFF2、ERβ的阳性表达率与术前化疗癌症组比较无显著性差异(均P>0.05)。子宫内膜癌组织中RECK、TFF2的阳性表达与临床分期、组织学分级、肌层浸润程度及淋巴结转移密切相关;ERβ的表达与组织学分级、肌层浸润程度及淋巴结转移密切相关,而与临床分期无相关性,子宫内膜癌组织存在p16基因全部或部分甲基化,正常子宫内膜组织不存在p16基因甲基化现象;其中子宫内膜癌组织中p16甲基化有42例(52.50%),p16甲基化与临床分期,组织学分级、肌层浸润与淋巴结转移密切相关。结论 RECK、TFF2、ERβ、p16甲基化在子宫内膜癌的不同组织学分级、肌层浸润程度与有无淋巴结转移中的表达各异,并在子宫内膜癌的诊治中具有临床应用价值。
基金Shaanxi Foundation for Innovation Team of Science and Technology(No.2018TD-003)Project from State Key Laboratory of Cancer Biology(No.CBSKL2019ZZ07)
文摘To the Editor:Gastric cancer(GC)ranks third in incidence and mortality both worldwide and in China.[1,2]Intestinal metaplasia(IM)significantly increases risk of GC so that identifying high-risk IM patients who will progress to GC is crucial.Currently,the effect of many risk-stratification methods for gastric precancerous lesions(GPLs)was minimal.Monoclonal gastric cancer 7 antigen(MG7-Ag)combined with cyclooxygenase-2 has been shown earlywarning value for the progression of GPLs.[3]The expression of human telomerase reverse transcriptase(hTERT)was proven to be related to state of cell proliferation in IM tissue.[4]Loss expression of trefoil factor family 2(TFF2)from spasmolytic polypeptideexpressing metaplasia to IM may lead to a hyperproliferation and deleterious mutations.[5]We tried to construct and verify a multimolecular prediction model included MG7-Ag and hTERT and TFF2,which could identify high-risk IM patients and have early-warning value for GC.
