Relationship between somatostatin receptor subtype expression and clinicopathology,Ki-67,Bcl-2 and p53 in colorectal cancer

AIM： To study the SSTR1, 2, 3, 4, 5 expression and their relationships with clinico-pathological factors, cell proliferation, Bcl-2 and p53 expression in colorectal cancer cells. METHODS： Immunohistochemical staining of five SSTR subtypes, Ki-67, Bcl-2 and p53 was performed by the standard streptavidin-peroxidase （SP） technique for the paraffin sections of 127 colorectal cancers, and expression of five SSTR subtypes in 40 specimens of normal colorectal mucosae was detected with the same method. RESULTS： Positive staining for five SSTR subtypes was observed in colorectal cancer cells and normal colorectal mucosae. SSTR1 was the most predominant subtype in both colorectal cancer and normal colorectal mucosa, and the second was SSTR5 or SSTR2. As compared with normal colorectal mucosa, SSTR4 was more frequently expressed in colorectal cancer cells （2.5% vs 18.9%, P〈 0.05）; the expression of SSTR2, 4, 5 in moderately to well differentiated colorectal adenocarcinoma was significantly higher than that in poorly differentiated ones （P〈 0.05）, the SSTR1 expression in colorectal cancer with positive lymph node metastasis was significantly higher than that with negative lymph node metastasis （72.2% and 54.5%, P〈 0.05）. In addition, in the ulcerative type of colorectal cancer, SSTR2 expression was obviously decreased （P 〈 0.05）; the correlation did not reach a statistical significance between the five SSTR subtypes expression and Dukes＇stages （P〉 0.05）, but the frequency of SSTR1 expression increased with Dukes＇ stage, while SSTR3 and SSTR5 expression decreased with Dukes＇ stage. Moreover, there was no correlation between expression of the five SSTR subtypes and other clinicopathological factors such as age, sex, tumor site, tumor depth, distant metastasis. The proliferative indexes in colorectal cancer cells with negative expression of SSTR2 and SSTR3 were significantly higher than that with positive expression （P〈0.05）. The Bcl-2 expression in colorectal cancer cells with positive expression of SSTR1, 2, 3, 5 was significantly lower than that with negative expression （P〈 0.05）. There was no correlation between five SSTR subtypes and p53 expression. CONCLUSION： The most predominant SSTR subtype is SSTR1, and the second is SSTR2 or SSTR5, Five SSTR subtypes play different roles in the development of colorectal cancer, SSTR2 and SSTR3 can inhibit the proliferation and promote apoptosis of tumor cells.

Long-term omeprazole and esomeprazole treatment does not significantly increase gastric epithelial cell proliferation and epithelial growth factor receptor expression and has no effect on apoptosis and p53 expression

AIM： To study the effect of proton pump inhibitor （PPI） treatment on patients with reflux esophagitis and its in vivo effect on apoptosis, p53- and epidermal growth factor receptor （EGFR） expression. METHODS： After informed consent was obtained, gastric biopsies of the antrum were taken from patients with reflux oesophagitis prior to and after 6 mo of 20 mg omeprazole （n = 24） or 40 mg esomeprazole （n = 22） therapy. Patients did not take any other medications known to affect the gastric mucosa. All patients were Helicobacter pylori negative as confirmed by rapid urease test and histology, respectively. Cell proliferation, apoptosis, EGFR, and p53 expression were measured by immunohistochemical techniques. At least 600 glandular epithelial cells were encountered and results were expressed as percentage of total cells counted. Was considered statistically significant. RESULTS： Although there was a trend towards increase of cell proliferation and EGFR expression both in omeprazole and esomeprazole treated group, the difference was not statistically significant. Neither apoptosis nor p53 expression was affected. CONCLUSION： Long-term PPI treatment does not significantly increase gastric epithelial cell proliferation and EGFR expression and has no effect on apoptosis and p53 expression.

Anticancer therapeutic strategies for targeting mutant p53-Y220C

The tumor suppressor p53 is a transcription factor with a powerful antitumor activity that is controlled by its negative regulator murine double minute 2(MDM2,also termed HDM2 in humans)through a feedback mechanism.At the same time,TP53 is the most frequently mutated gene in human cancers.Mutant p53 proteins lose wild-type p53 tumor suppression functions but acquire new oncogenic properties,among which are deregulating cell proliferation,increasing chemoresistance,disrupting tissue architecture,and promoting migration,invasion and metastasis as well as several other pro-oncogenic activities.The oncogenic p53 mutation Y220C creates an extended surface crevice in the DNA-binding domain destabilizing p53 and causing its denaturation and aggregation.This cavity accommodates stabilizing small molecules that have therapeutic values.The development of suitable small-molecule stabilizers is one of the therapeutic strategies for reactivating the Y220C mutant protein.In this review,we summarize approaches that target p53-Y220C,including reactivating this mutation with small molecules that bind Y220C to the hydrophobic pocket and developing immunotherapies as the goal for the near future,which target tumor cells that express the p53-Y220C neoantigen.

抑癌基因p53诱导Wnt通路抑制因子sFRP的表达

目的研究p53对Wnt通路抑制因子sFRP表达的调节作用。方法将携带p53基因的复制缺陷型腺病毒载体(Adp53)导入到p53缺失的人肝癌细胞株Hep3B中,以流式细胞术检测Adp53转基因情况,以RT-PCR技术检测p53对sFRP表达的调节作用。结果sFRP mRNA水平在转染p53 20 h后即有明显升高,其中以32 h达最高水平,随后逐渐降低。量效关系研究表明在转染剂量为0.05、0.5、5、50 pfu/cell的Adp53 sFRP mRNA表达均有显著增高,尤以5 pfu/cell时表达水平最高。结论p53能明显诱导Wnt通路抑制剂sFRP的mRNA表达。

Transgelin induces apoptosis of human prostate LNCaP cells through its interaction with p53

The androgen receptor （AR） and its coregulators have important roles in the carcinogenesis of prostate cancer.p53 is an important tumour suppressor gerte,and the absence of a fundamental p53 response may predispose to cancer. Transgelin,known as an ARA54-associated AR inhibitor,can suppress AR function in LNCaP cells. In addition to these effects,we aimed to elucidate the proapoptotic effects of the protein on LNCaP and its underlying mechanisms,especially the interaction between transgelin and p53. Cell counting,flow cytometric analysis and terminal deoxynucleotidyl transferase-dUTP nick-end labelling assays were applied to measure the proapoptotic effect of transgelin. Using western blotting of p53 and double immunofluorescence staining of p53 with transgelin,we show that transfection of transgelin results in increasing cytoplasmic translocation of p53 and upregulation of p53 expression. We also found an interaction between transgelin and p53 in vivo by mammalian two-hybrid and coimmunoprecipitation assays. The activation of the mitochondria-associated apoptosis pathway was observed in LNCaP cells after transfection with transgelin. These results are indicative of p53-mediated mitochondria-associated apoptotic effects of transgelin on LNCaP cells in addition to its known suppressive effects on the AR pathway.

Evaluation of malignancy using Ki-67,p53,EGFR and COX-2 expressions in gastrointestinal stromal tumors

AIM:To investigate the role of expressions of Ki-67, p53,epidermal growth factor receptor(EGFR)and cyclooxygenase-2(COX-2)in gastrointestinal stromal tumor(GIST)grading and prognosis. METHODS:Tumor tissue was collected retrospectively from 96 patients with GIST.Antibodies against Ki-67, p53,EGFR and COX-2 were used for immunohistochemical staining.Tumor grading was designated according to a consensus system and the staining was quantified in 3 categories for each antibody in the statistical analysis. RESULTS:The Ki-67 expression in GISTs was significantly associated with the size of the tumors,mitotic rate and the risk of malignancy(x2=15.51,P=0.02; x2=22.27,P<0.001;x2=20.05;P<0.001).The p53 expression was also significantly correlated with mitotic rate and the risk of malignancy(x2=9.92,P= 0.04;x2=9.97;P=0.04).Over-expression of Ki-67 was strongly correlated with poor survival(x2=10.44, P=0.006),but no correlation was found between the expression of p53,EGFR or COX-2 and survival. Multivariate analysis further demonstrated that Ki-67 expression(relative risk=15.78,95%CI:4.25-59.37) could be used as an independent prognostic value for GIST patients.Adjuvant imatinib therapy could improve clinical outcomes in the patients with high risk and intermediate risk of recurrence after complete tumor resections(median survival time:52 mo vs 37 mo, x2=7.618,P=0.006). CONCLUSION:Our results indicated that the expression of Ki-67 could be used as an independent prognostic factor for GIST patients.

Estradiol agonists inhibit human Lo Vo colorectal-cancer cell proliferation and migration through p53

AIM: To investigate the effects of 17&#x003b2;-estradiol via estrogen receptors (ER) or direct administration of ER agonists on human colorectal cancer.

Clinicopathological characteristics of human epidermal growth factor receptor 2-positive Barrett's adenocarcinoma

AIM:To compare the clinicopathological characteristics of human epidermal growth factor receptor 2 (HER2)-positive and HER2-negative Barrett's adenocarcinoma in Japan. METHODS:We performed immunohistochemical analysis of HER2 in 30 samples taken from patients with Barrett's adenocarcinoma and dual color in situ hybridization in cases showing 2+ reactions. We compared the clinicopathological characteristics of HER2-positive and HER2-negative patients.RESULTS:HER2 positivity was identified in 8 (27%) carcinoma samples. We found that HER2 expression was associated with p53 overexpression (100% vs 52.6% in pT1 tumor; 100% vs 54.5% in all stage tumor, P < 0.05) and protruding lesions at the early disease stage. There was no association between the mucin phenotype of the carcinomas and prognosis. HER2 expression and low clinical stage were unexpectedly different between Barrett's adenocarcinoma patients and gastric cancer patients, but the macroscopic features may be associated with earlier diagnosis in these patients. CONCLUSION:Our results suggest that HER2-positive Barrett's adenocarcinomas are associated with p53 overexpression and lesion protrusion at the early disease stage.

流感病毒感染p53+/+和p53-/-小鼠肺组织中TLR及其相关基因表达研究

目的分析p53介导的抗流感病毒感染过程中,Toll-like receptors(TLRs)及其信号通路分子mRNA表达变化,为p53抗病毒作用机制的解析提供帮助。方法流感病毒PR8感染p53+/+、p53-/-小鼠,通过小鼠表达谱芯片分析病毒感染后小鼠肺脏中TLR家族成员及其信号通路分子基因表达差异。结果流感病毒感染后,表达谱芯片分析结果表明,p53+/+小鼠肺组织中TLR8、TLR9以及信号通路中的NF-κB、TNF-α、IFN-α表达等基因的表达与p53-/-小鼠相比差异显著。结论 TLRs及其信号通路分子参与了p53介导的抗流感病毒作用。

Human umbilical cord mesenchymal stem cells attenuate diabetic nephropathy through the IGF1R-CHK2-p53 signalling axis in male rats with type 2 diabetes mellitus

diabetes mellitus(DM)is a disease syndrome characterized by chronic hyperglycaemia.A long-term high-glucose environment leads to reactive oxygen species(ROS)production and nuclear DNA damage.human umbilical cord mesenchymal stem cell(HUcMSC)infusion induces significant antidiabetic effects in type 2 diabetes mellitus(T2DM)rats.Insulin-like growth factor 1(IGF1)receptor(IGF1R)is important in promoting glucose metabolism in diabetes;however,the mechanism by which HUcMSC can treat diabetes through IGF1R and DNA damage repair remains unclear.In this study,a DM rat model was induced with high-fat diet feeding and streptozotocin(STZ)administration and rats were infused four times with HUcMSC.Blood glucose,interleukin-6(IL-6),IL-10,glomerular basement membrane,and renal function were examined.Proteins that interacted with IGF1R were determined through coimmunoprecipitation assays.The expression of IGF1R,phosphorylated checkpoint kinase 2(p-CHK2),and phosphorylated protein 53(p-p53)was examined using immunohistochemistry(IHC)and western blot analysis.Enzyme-linked immunosorbent assay(ELISA)was used to determine the serum levels of 8-hydroxydeoxyguanosine(8-OHdG).Flow cytometry experiments were used to detect the surface markers of HUcMSC.The identification of the morphology and phenotype of HUcMSC was performed by way of oil red“O”staining and Alizarin red staining.DM rats exhibited abnormal blood glucose and IL-6/10 levels and renal function changes in the glomerular basement membrane,increased the expression of IGF1 and IGF1R.IGF1R interacted with CHK2,and the expression of p-CHK2 was significantly decreased in IGF1R-knockdown cells.When cisplatin was used to induce DNA damage,the expression of p-CHK2 was higher than that in the IGF1R-knockdown group without cisplatin treatment.HUcMSC infusion ameliorated abnormalities and preserved kidney structure and function in DM rats.The expression of IGF1,IGF1R,p-CHK2,and p-p53,and the level of 8-OHdG in the DM group increased significantly compared with those in the control group,and decreased after HUcMSC treatment.Our results suggested that IGF1R could interact with CHK2 and mediate DNA damage.HUcMSC infusion protected against kidney injury in DM rats.The underlying mechanisms may include HUcMSC-mediated enhancement of diabetes treatment via the IGF1R-CHK2-p53 signalling pathway.

EGFR、p53与Ki-67在国人胶质瘤中表达研究

表皮生长因子受体（epidermal grotwth factor receptor，EGFR）的过表达或扩增、激活以及抑癌基因p53的缺失、突变失活均是导致胶质瘤的主要分子事件。本文应用免疫组化方法对50例国人脑胶质瘤中表皮生长因子受体EGFR、p53蛋白及胶质瘤恶性程度的标志Ki-67标记指数（Ki-67 LI）联合检测，对胶质瘤分子生物学行为进行探讨，为基因治疗在胶质瘤中的应用提供理论依据。

人子宫平滑肌肿瘤的雌、孕激素受体和p^(53)蛋白表达

目的：探讨子宫平滑肌瘤与雌、孕激素受体的关系；了解ｐ５３蛋白在不同组织学类型肌瘤细胞内的表达情况。方法：直接荧光组织化学法和免疫组化法。结果：子宫肌瘤雌、孕激素受体阳性率为６５．５２％，高于子宫肌壁的雌、孕激素受体３６．３６％的阳性率。两组差异有显著性（Ｐ＜０．０５）。６０例子宫平滑肌瘤ｐ５３蛋白总阳性表达率为１８．８３％，良性平滑肌瘤组、富细胞型及子宫肉瘤组ｐ５３蛋白阳性率分别为１３．３３％、１５％和４０％。肉瘤组ｐ５３蛋白阳性率明显高于良性肌瘤组，但差异无显著性。结论：雌、孕激素对子宫平滑肌瘤的发生均有一定作用。人子宫平滑肌瘤ｐ５３蛋白阳性表达率低于女性生殖道上皮源性肿瘤，而与人纤维源性肿瘤的ｐ５３蛋白阳性表达率接近。

20例原发性输卵管癌组织中ER、PR与p^(53)蛋白的表达

目的：分析原发性输卵管腺癌的ＥＲ、ＰＲ及ｐ５３蛋白的表达，研究其与该肿瘤的临床分期、病理分级及患者预后的关系。方法：材料选自２０份原发性输卵管腺癌及１０份正常输卵管组织的存档石蜡包埋标本，采用免疫组化法检测。结果：在２０份原发性输卵管癌标本中，ＥＲ、ＰＲ的阳性表达率分别为２５％和１５％，稍高于正常输卵管组的１０％，但差异无显著性（Ｐ＞０．０５）；ｐ５３蛋白在癌症组的阳性表达率为４０％，对照组则无１例阳性表达，差异有显著性（Ｐ＜０．０５）；ｐ５３蛋白在晚期、分化差及预后不良的输卵管癌病例中的表达呈上升趋势，但未达到统计学意义（Ｐ＞０．０５）；对侧输卵管炎症的存在与ｐ５３蛋白的阳性表达呈负相关（Ｐ＜０．０５）。结论：在原发性输卵管癌中，ＥＲ、ＰＲ有一定程度的表达，但均较低；ｐ５３基因的突变可能参与了该肿瘤的发生，并可作为综合判断其恶性程度及患者预后的指标之一。

有和无内分泌分化大肠癌P^(53)和表皮生长因子受体表达的比较研究

目的：研究大肠癌内分泌分化与P53表皮生长因子受体（EGFR）表达的关系及意义。方法：采用免疫组织化学ABC法，检测79例大肠癌手术切除组织中嗜铬蛋白A（CGA），P53及EGFR的表达变化。结果：大肠癌组织中CGA，P53及EGFR的表达率分别为：35.4％（28／79〕，60．8％(48／79）和30．4％（24／79），有内分泌分化的大肠癌P53表达率及其免疫阳性细胞数分别为78．6％（22／28）和984．3士702．0／mm2，无内分泌分化者分别为51．0％和589．5士489．5／mm2，前者明显高于后者（P<O．025；P＜0．05）。有和无内分泌分化的大肠癌EGFR的表达率分别为46．4％（13／28）和21．6％（11／51），差异有显著性（P＜0．025）。结论：有内分泌分化的大肠癌P53及EGFR的表达明显高于无内分泌分化者。

Emodin induces apoptosis in human prostate cancer cell LNCaP

Aim： To elucidate effects and mechanisms of emodin in prostate cancer cells. Methods： Viability of emodin-treated LNCaP cells and PC-3 cells was measured by MTT assay. Following emodin treatments, DNA fragmentation was assayed by agarose gel electrophoresis. Apoptosis rate and the expression of Fas and FasL were assayed by flow cytometric analysis. The mRNA expression levels of androgen receptor （AR）, prostate-specific antigen （PSA）, p53, p21, Bcl-2, Bax, caspase-3, -8, -9 and Fas were detected by RT-PCR, and the protein expression levels of AR, p53 and p21 were detected by Western blot analysis. Results： In contrast to PC-3, emodin caused a marked increase in apoptosis and a decrease in cell proliferation in LNCaP cells. The expression of AR and PSA was decreased and the expression of p53 and p21 was increased as the emodin concentrations were increased. In the same time, emodin induced apoptosis of LNCaP cells through the upregulation of caspase-3 and -9, as well as the increase of Bax/Bcl-2 ratio. However, it did not involve modulation of Fas or caspase-8 protein expression. Conclusion： In prostate cancer cell line, LNCaP, emodin inhibites the proliferation by AR and p53-p21 pathways, and induces apoptosis via the mitochondrial pathway.

子宫内膜癌雌孕激素受体及P^(53)表达的临床意义

检测子宫内膜癌组织中雌激素受体 (ER)、孕激素受体 (PR)及 P5 3表达的阳性率并探讨其临床意义。方法 :用免疫组化法对 33例子宫内膜癌标本进行了 ER、 PR及 P5 3 的检测。结果 :子宫内膜癌组织中 ER、 PR及 P5 3 的总阳性率分别为6 6 .5 7%、 6 3.6 4%、 2 7.2 7% ,ER、PR、P5 3阳性表达率与癌组织的细胞分化程度有关 ,随着子宫内膜癌组织学分级的增高 ,ER、 PR阳性表达率逐渐降低 ,而 P5 3 的阳性率逐渐升高 (P <0 .0 5 )。与临床分期关系无统计学意义。结论 :ER、 PR、 P5 3 及组织学分级均反映了子宫内膜癌的生物学行为 ,它们的测定对估计患者预后和临床选择内分泌治疗具有重要意义。

乳腺癌P_(53)抗癌基因雌激素受体孕激素受体的表达及其对治疗和预后的意义

应用免疫组织化学ＬＳＡＢ法，对４０例乳腺癌的Ｐ５３抗癌基因（Ｐ５３）、雌激素受体（ＥＲ）、孕激素受体（ＰＲ）进行了检测，结果显示：Ｐ５３阳性２４例（６０％），ＥＲ阳性２０例（５０％），ＰＲ阳性１５例（３７．５％），其中三项均阳性者７例，而Ｐ５３与ＥＲ均阳性者１４例，Ｐ５３与ＰＲ均阳性者１１例。结果表明：乳腺癌中Ｐ５３阳性率高于良性乳腺肿瘤，差异显著（Ｐ＜０．０１），ＥＲ阳性率随患者年龄增大而升高；５０岁以上患者阳性率明显高于５０岁以下者；而Ｐ５３、ＰＲ的表达与年龄无关。本组研究表明，ＥＲ、ＰＲ阳性患者采用手术治疗和术后放疗、化疗再增加内分泌治疗，可使患者５年、１０年生存率明显提高，而Ｐ５３阳性、ＥＲ阴性的患者无明显疗效。

鼻咽癌P^(53),C-myc,EGFR基因表达的研究

目的：Ｐ５３，Ｃ－ｍｙｃ及ＥＧＦＲ基因在鼻咽癌（ＮＰＣ）的发生、发展中可能起重要作用，基因的研究结果可能为ＮＰＣ的预防及诊疗提供科学的依据。方法：用免疫组化Ｓ－Ｐ法检测了３０例ＮＰＣ组织、１０例无瘤鼻咽部（ＮＰ）组织的Ｐ５３，Ｃ－ｍｙｃ及ＥＧＦＲ基因的异常表达情况。结果：①三种基因在ＮＰＣ组织中均有较高水平的表达，Ｐ５３蛋白阳性率为８６．７％（２６／３０），Ｃ－ｍｙｃ和ＥＧＦＲ蛋白阳性率均为７０％（２１／３０）；相应对照的无瘤ＮＰ组织的阳性率分别为０．０％（０／１０）、１０％（１／１０）和３０％（３／１０）。ＮＰＣ组织与无瘤ＮＰ组织的Ｐ５３，Ｃ－ｍｙｃ及ＥＧＦＲ蛋白阳性率高于无瘤ＮＰ组织相应的阳性率，差异有统计学意义（Ｐ＝０．００００、Ｐ＝０．００１３及Ｐ＝０．０３１８）；②ＮＰＣ按ＴＮＭ临床分期，将其Ｔ、Ｎ级数分别与Ｐ５３、Ｃ－ｍｙｃ，ＥＧＦＲ蛋白染色阳性程度进行相关分析，皆无统计学意义（Ｐ＞０．０５）；③Ｃ－ｍｙｃ与ＥＧＦＲ的阳性表达之间有相关性（Ｐ＝０．０００９）。结论：①Ｐ５３，Ｃ－ｍｙｃ，ＥＧＦＲ三种基因的高表达，说明其与ＮＰＣ的发生有一定关系，认为Ｐ５３的免疫组化检测可作为ＮＰＣ的辅助诊断依据；?

卵巢克鲁根勃瘤及卵巢癌P^(53)ER PR表达的研究

应用免疫组织化学BSA法检测32例卵巢克鲁根勃瘤、20例卵巢癌中P53蛋白、雌激素受体（ER）、孕激素受体（PR），其阳性表达率克鲁根勃瘤分别10.9％（13／32）、21.9％（7／32）、21．9％（7／32），卵巢癌依次为45％（9／20）、65％（13／20）、50％（10／20）。结果表明P53在两者中阳性表达率无显著相关性，ER、PR在高分化卵巢癌中阳性表达率明显高于克鲁根勃瘤及低分化卵巢癌（P＜0．001）。在卵巢癌中ER、PR阳性表达率与组织学分化程度正相关（P＜0．05）与预后负相关（P＜0．01）。

Histological Grading in Ductal Carcinoma in Situ of the Breast

Objective: To study the significance of histological grading as a prognostic factor in ductal carcinoma in situ of the breast. Methods: According to the Van Nuy’s classification, 32 cases of ductal carcinoma in situ (DCIS) of the breast were divided into three groups. Results: Low grade (well differentiated, low grade DCIS) 12 patients (37.5%); Intermediate grade, 9 patients (28.1%); High grade (poorly differentiated DCIS) 11 patients (34.4%). Among the high grade DCIS, the histologic subtypes were comedo (9 patients), micropapillary (1 patient) and solid (1 patient). The positive expression of c-erbB-2, p53 and MIB-1 in high grade DCIS was higher than that in intermediate and low grade DCIS. The difference between high grade and low grade DCIS was significant (p<0.05). The expression of ER in high grade DCIS was lower than that in intermediate and low grade DCIS. Conclusions: Histological grading of breast ductal carcinoma in situ may be a good prognostic factor.