HSPA5 CD44v6在子宫内膜癌的表达及临床价值

目的探讨热休克蛋白5(HSPA5)和CD44v6在子宫内膜癌中的表达及临床价值。方法选取2018年1月至2020年12月上海市长宁区妇幼保健院手术切除的93例子宫内膜癌标本,用免疫组织化学方法作HSPA5和CD44v6的表达检测,结合其临床病理指标,进行统计学分析。结果在93例子宫内膜癌中,HSPA5、CD44v6表达的阳性率分别为71.0%(66/93)和65.6%(61/93)。HSPA5在子宫内膜癌的不同病理分期、是否绝经及肿瘤浸润子宫壁深度不同(浸润子宫壁深度<1/2组;浸润子宫壁深度≥1/2组)的表达结果差异无统计学意义(均为P>0.05);而HSPA5在子宫内膜癌的不同组织学类型、淋巴结有无转移组的表达结果差异均有统计学意义(P<0.01,P<0.005)。CD44v6在子宫内膜癌的不同临床分期、是否绝经及不同组织学类型的表达结果差异无统计学意义(均为P>0.05);而CD44v6在肿瘤浸润子宫壁不同深度(浸润子宫壁深度<1/2组;浸润子宫壁深度≥1/2组)、淋巴结有无转移组的表达结果差异均有统计学意义(P<0.001,P<0.005)。结论HSPA5、CD44v6在子宫内膜癌中存在高表达,二者与子宫内膜癌的生物学行为有关,在子宫内膜癌的发生、发展中有着重要的调控作用。同步进行HSPA5、CD44v6在子宫内膜癌的表达检测,在判断其预后和指导其临床治疗上有重要价值。

CD133 和 CD44 在胃腺癌中的表达及其与 E-cadherin 的关系

目的探讨原发性胃腺癌(gastric adenocarcinoma,GAC)组织中CD133、CD44和E-cadherin蛋白表达的关系及临床病理意义。方法应用免疫组织化学ElivisionTMplus法检测145例胃腺癌组织和30例正常胃黏膜组织中CD133、CD44和E-cadherin蛋白的表达。结果在正常胃黏膜组织中,CD133、CD44和E-cadherin蛋白的阳性表达率分别为10.0%、3.3%和100%;在GAC组中,CD133、CD44和E-cadherin蛋白的阳性表达率分别为46.9%、47.6%和42.8%,两组之间,差异均有显著性(P<0.05)。CD133、CD44和E-cadherin蛋白的表达水平均与肿瘤的大小、分化程度、淋巴结转移与否、浸润深度、临床分期及术后生存期有关(全部P<0.05);CD133的表达与CD44的表达呈正相关(P<0.05),CD133和CD44的表达均与E-cadherin的表达呈负相关(P<0.005)。Kaplan-Meier生存分析表明,CD133和CD44蛋白表达阳性的患者其生存率明显低于其阴性表达患者(P<0.001);Ecadherin蛋白表达阳性的患者其生存率明显高于其阴性表达患者。Cox回归分析:CD133、CD44和E-cadherin的表达是影响GAC患者术后生存率的独立预后因素。结论 CD133、CD44和E-cadherin的表达水平在GAC组织中与肿瘤组织的大小、分化程度、临床分期、浸润深度、淋巴结转移与否及预后等均有关;在GAC组织中联合检测CD133、CD44和E-cadherin对判断预后有价值。

hTERT CD44v6在大肠癌中的表达及意义

目的:探讨hTERT、CD44v6的表达与大肠癌发生、浸润转移及预后的关系。方法:对大肠腺瘤组织及大肠癌组织进行免疫组织化学染色,分析hTERT及CD44v6在大肠癌中表达的相关性。结果:hTERT及CD44v6在大肠癌组织中的阳性表达率分别为70.73%、53.66%,明显高于大肠腺瘤组织中的10.53%、10.53%(P<0.01);hTERT及CD44v6在有淋巴结转移的大肠癌患者的阳性表达率分别为93.33%、86.67%,明显高于无淋巴转移患者的阳性表达率53.85%、38.46%(P<0.01);hTERT及CD44v6在TNMⅢ、Ⅳ期大肠癌患者的阳性表达率分别为94.12%、82.35%,明显高于TNMⅠ、Ⅱ期的54.17%、33_33%(P<0.01);在生存期<5年大肠癌中的阳性表达率分别为90.91%、77.27%,明显高于生存期≥5年的47.37%、26.32%(P<0.01);hTERT和CD44v6在性别、年龄、肿物大小、肿物位置及浸润深度中的阳性表达差异均无统计学意义(P>0.05)。结论:hTERT和CD44v6在大肠癌组织中具有高表达,特别是在有淋巴结转移、TNMⅢ、Ⅳ期和生存期<5年组中有较高表达,说明两者可能在大肠癌的发生、浸润转移过程中起着重要作用,并且与大肠癌患者的预后有关;联合检测hTERT和CD44v6可以作为大肠癌浸润转移的判断和预后评估的参考指标。

p53 CD44V6及nm23/NDPK在非小细胞肺癌中表达的相关性分析

目的:探讨非小细胞肺癌(NSCLC)中p53、CD44V6及nm23/NDPK表达的相关性及预后意义。方法:采用免疫组织化学SP法对69例NSCLC组织中p53、CD44V6及nm23/NDPK进行检测。结果:p53、CD44V6及nm23/NDPK的表达与NSCLC患者的临床分期(P<0.05,P<0.01)、淋巴结转移(P<0.05,P<0.01)及生存率(P<0.05)密切相关。p53、CD44V6表达与nm23/NDPK在NSCLC中的表达呈明显负相关(P<0.01)。结论:联合检测NSCLC中p53、CD44V6及nm23/NDPK的表达对于判断预后和指导治疗可能具有实用价值。

Bcl-2 Survivin CD44v6的表达HPV_(16/18)感染及其与宫颈癌发生的相关性探讨

目的:研究Bcl-2、Survivin、CD44v6在宫颈癌的表达及其与HPV16/18感染的相关性,以探讨宫颈癌的发生机制。方法:采用免疫组化对宫颈癌、上皮内瘤样病变(CIN)和正常宫颈进行Bcl-2、Survivin、CD44v6的检测,同时采用PCR检测HPV16/18感染情况。结果:Bcl-2、Survivin、CD44v6的阳性率在宫颈癌与CIN、正常宫颈间有显著差异,且随着临床分期、病理分级的升高阳性率差异有显著性,其阳性率与淋巴结转移有关。HPV16/18在正常宫颈组织、CIN和宫颈癌中的阳性率逐渐升高,差异有显著性。Bcl-2和Survivin均与HPV16/18感染有相关性。结论:宫颈癌中Bcl-2、Survivin、CD44v6的异常表达和HPV16/18感染有关,与宫颈癌的恶变程度有关,可作为宫颈癌筛查预后判断的有利指标。

VEGF CD44v6 MMP-9在子宫内膜异位症组织中的表达及临床意义

目的探讨血管内皮生长因子(vascular endothelial growth factor,VEGF)、黏附分子CD44v6、基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)在子宫内膜异位症发病机制中的作用及临床意义。方法通过免疫组化S-P法分别对60例卵巢异位内膜、52例子宫内膜异位症在位子宫内膜和44例正常子宫内膜组织分别行VEGF、CD44v6和MMP-9的检测,并进行观察和对比。结果 VEGF、CD44v6、MMP-9在卵巢异位内膜中的表达与在位内膜组和正常内膜组比较,有明显增强(P<0.01);在位内膜与正常子宫内膜相比,VEGF在位内膜中的表达高于正常内膜中的表达,有统计学意义(P<0.01),CD44v6及MMP-9在在位内膜中的表达略高于正常内膜中的表达,但无统计学意义(P>0.05)。结论 VEGF、CD44v6、MMP-9在卵巢异位内膜中呈高表达状态,异位内膜组VEGF和CD44v6之间呈正相关(r1=0.462,χ2=42.038,P=0.000);MMP-9与CD44v6之间呈正相关(r2=0.558,χ2=70.534,P=0.000);MMP-9与VEGF呈正相关(r3=0.397,χ2=29.187,P=0.000),表明与异位内膜的高侵袭性和高血管新生能力有关,可能在EMS的发生发展中起重要作用。

KAI1 CD44v3表达与喉癌的相关性研究

目的了解喉癌组织中KAI1和CD44v3表达的情况,探讨其在喉癌中的意义及与淋巴结转移的关系。方法采用免疫组织化学(SP)法分别检测40例喉癌患者的咽喉黏膜组织、30例正常体检者咽喉黏膜组织中KAI1和CD44v3的表达水平。并分析两者在喉癌不同分化程度、临床分期、有无淋巴结转移的癌组织中表达的差异及其相关性。结果癌组织中KAI1的表达低于在正常咽喉黏膜组织中的表达(χ2=8.86,P<0.01),在癌组织中表达与分化程度、临床分期及是否有淋巴结转移无关(P>0.05),但阳性表达的强度与临床分期及是否有淋巴结转移有关(P<0.05);CD44v3在癌组织中的表达高于正常咽喉黏膜组织(χ2=18.36,P<0.01),表达与临床分期及是否有淋巴结转移有关(P<0.05),与分化程度无关。KAI1及CD44v3在癌组织中的表达具有相关性(χ2cs=4.19,P<0.05),关联系数r=0.67。结论KAI1的表达可能抑制喉癌的发生及转移过程,CD44v3可能促进了喉癌的发生和转移。

Construction of cell lines with CD44 cDNA and its application in hepatocellular carcinoma

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原代口腔鳞癌细胞的分离培养及CD133 CD44的表达

目的探讨从人新鲜口腔鳞癌组织中培养原代鳞癌细胞的方法以及干细胞表面标志物CD133、CD44表达量的检测情况。方法选取该院2016~2017年行口腔鳞状细胞癌切除术的新鲜肿瘤标本6例,应用机械法获得原代口腔鳞癌细胞并用酶消化法传代。应用细胞形态学及角蛋白免疫组化方法对原代鳞癌细胞进行鉴定。流式细胞仪技术检测鳞癌细胞中干细胞表面标志物CD133、CD44的表达情况。结果(1)成功培养出纯化的口腔鳞癌细胞,并能成功传代,至目前为止已传至27代。(2)培养出的OSCCs呈多边形铺路石状,角蛋白染色其胞浆中可见阳性棕色染色。(3)流式细胞检测鳞癌细胞中干细胞表面标志物CD133的平均表达率为0.41%、CD44的平均表达率为33.76%。结论从人新鲜口腔鳞癌标本中利用机械法可成功获得大量原代口腔鳞癌细胞并能顺利传代,口腔鳞癌细胞中干细胞表面标志物CD44表达偏高,CD133的表达量明显偏低。

Expressions of osteopontin and CD44v6 and their clinical significance in pituitary adenomas

Objective To explore expressions of osteopontin (OPN) and CD44v6 in pituitary adenomas and to evaluate relationship between expressions and invasiveness of tumor. Methods Immunohistochemistry was used to detect expressions of OPN and CD44v6 in 80 pituitary adenomas,and

Expression and significance of CD44s, CD44v6, and nm23 mRNA in human cancer

AIM: To investigate the relationship between the expression levels of nm23 mRNA, CD44s, and CD44v6,and oncogenesis, development and metastasis of human gastric adenocarcinoma, colorectal adenocarcinoma,intraductal carcinoma of breast, and lung cancer.METHODS: Using tissue microarray by immuhistochemical (IHC) staining and in situ hybri-dization (ISH), we examined the expression levels of nm23mRNA, CD44s, and CD44v6 in 62 specimens of human gastric adenocarcinoma and 62 specimens of colorectal adenocarcinoma; the expression of CD44s and CD44v6in 120 specimens of intraductal carcinoma of breast and 20 specimens of normal breast tissue; the expression of nm23 mRNA in 72 specimens of human lung cancer and 23 specimens of normal tissue adjacent to cancer.RESULTS: The expression of nm23 mRNA in the tissues of gastric and colorectal adenocarcinoma was not significantly different from that in the normal tissues adjacent to cancer (P＞0.05), and was not associated with the invasion of tumor and the pathology grade of adenocarcinoma (P＞0.05). However, the expression of nm23 mRNA was correlated negatively to the lymph node metastasis of gastric and colorectal adenocarcinoma (r = -0.49, P＜0.01; r = -4.93, P＜0.01). The expression of CD44s in the tissues of gastric and colorectal adenocarcinoma was significantly different from that in the normal tissues adjacent to cancer (P＜0.05;P＜0.01). CD44v6 was expressed in the tissues of gastric and colorectal adenocarcinoma only, the expression of CD44v6 was significantly associated with the lymph node metastasis, invasion and pathological grade of the tumor (r = 0.47, P＜0.01; r = 5.04, P＜0.01). CD44sand CD44v6 were expressed in intraductal carcinoma of breast, the expression of CD44s and CD44v6 was significantly associated with lymph node metastases and invasion (P＜0.01). However, neither of them was expressed in the normal breast tissue. In addition, the expression of CD44v6 was closely related to the degree of cell differentiation of intraductal carcinoma of breast (x2= 5.68, P＜0.05). The expressional level of nm23mRNA was closely related to the degree of cell differentiation (P＜0.05) and lymph node metastasis (P＜0.01), but the expression of nm23 gene was not related to sex, age, and type of histological classification (P＞0.05).CONCLUSION: Patients with overexpression of CD44s and CD44v6 and low expression of nm23 mRNA have a higher lymph node metastatic rate and invasion. In addition, overexpression of CD44v6 is closely related to the degree of cell differentiation. Detection of the three genes is able to provide a reliable index to evaluate the invasion and metastasis of tumor cells.

Expression of human chorionic gonadotropin, CD44v6 and CD44v4/5 in esophageal squamous cell carcinoma

AIM: To study the relationship between the expression of human chorionic gonadotropin (HCG), CD44v6,CD44v4/5 and the infiltration, metastasis of esophageal squamous cell carcinoma. METHODS: By labeled streptavidin-biotin technique,the expressions of HCG, CD44v6, and CD44v4/5 in 42 patients with esophageal squamous cell carcinoma were examined.RESULTS: The positive rate of HCG expression in patients with lymph node metastasis was 85.71%(18/21), higher than that (57.14%, 12/21) in those without lymph node metastasis (P＜0.05). The positiverate of CD44v6 expression was 71.43% (15/21) in lymph node metastasis group, and 38.09% (8/21) in nonmetastasis group; there was a significant difference between the two groups (P＜0.05). The positive rateof CD44v4/5 expression was 76.19% (16/21) inlymph node metastasis group, and 42.86% (9/21) in non-metastasis group; there was also a significant difference between them (P＜0.05). From grade I to grade Ⅲ in differentiation, the positive rate of HCG expression was 84.62% (11/13), 70.59% (12/17) and 58.33% (7/12), respectively; there was no significant difference among them (P＞0.05). The positive rate of CD44v6 expression in grades Ⅰ -Ⅲ of cancer tissues was 76.92% (10/13), 52.94% (9/17), and 33.33%(4/12) respectively; there was no significant difference among them. The positive rate of CD44v4/5 expression in grades Ⅰ-Ⅲ of cancer tissues was 69.23% (9/13),64.71% (11/17), and 41.67% (5/12) respectively; there was no significant difference among the three groups. There was no correlation between the positive rates ofHCG and CD44v6, CD44v4/5 expression. Cancer cells in carcinomatous emboli and those infiltrating into vascular wall strongly expressed HCG, CD44v6, and CD44v4/5. CONCLUSION: Expression of HCG, CD44v6, and CD44v4/5 in esophageal squamous cell carcinoma is related to its infiltration and metastasis. HCG, CD44v6, and CD44v4/5 have different effects on the infiltration and metastasis of esophageal squamous cell carcinoma.

The expression and clinical significance of CD44v in human gastric cancers

INTRODUCTIONCD44 was originally implicated as a'homing'receptor directing the migration of recirculatinglymphocytes.CD44 expression has beenconfirmed not only in lymphocytes but also in awide variety of epithelial tissues.It is considered asan important cell adhesion molecule for cell to cellinteractions.Molecular cloning and analysis

Role of VEGF and CD44v6 in differentiating benign from malignant ascites

AIM: To detect the vascular endothelial growth factor (VEGF)and soluble splice variant 6 of CD44 (sCD44v6) levels in ascites and to explore their role in differentiating benign from malignant ascites.METHODS: Cirrhotic ascites (n=36), tuberculosis ascites (n=8) and malignant ascites (n=23) were collected and studied. Concentrations of soluble VEGF and sCD44v6 in various kinds of ascites (n=67) were measured using a sandwich enzyme-linked immunoadsorbent assay.RESULTS: VEGF and sCD44v6 levels in malignant ascites were 640.74±264.81 pg/ml and 89.22±38.20 ng/ml,respectively, both of which were significantly higher than those in cirrhotic ascites and tuberculous ascites (q=18.98,11.89 and q=8.92, 5.09; P<0.01). However, the levels of VEGF and sCD44v6 in cirrhotic and tuberculous ascites had no significant difference (q=0.48, 0.75; P>0.05).Furthermore, VEGF levels in malignant ascites in patients with ovarian cancer were higher than those with gastric and colon cancer (q=5.03, 6.79; P<0.01, respectively). But differences of VEGF levels between gastric and colon cancer were not significant (q=1.90, P>0.05). Whereas, sCD44v6 levels in malignant ascites from patients with ovadan, gastric and colon cancer had no significant difference (q=0.06, 0.91,0.35;P>0.05, respectirely). In comparison with cirrhotic and tuberculous ascites, when the upper limit of its VEGF mean levels 119.44 pg/ml (70.90±48.54) and sCD44v6 mean levels 63.59 ng/ml (44.42±19.17) was taken as the minimum cutoff limit, the sensitivity and specificity of VEGF and sCD44v6 of this assay to the diagnos is of malignant ascites were 91.3 %, 90.9 % and 73.9 %, 88.7 % respectively.CONCLUSION: Elevated levels of VEGF and sCD44v6 may be useful in differential diagnosis of benign and malignant ascites.

Micro RNA-145 exerts tumor-suppressive and chemoresistance lowering effects by targeting CD44 in gastric cancer

AIM To determine the potential roles of CD4 and micro RNA(mi R)-145 in gastric cancer.METHODS The levels of CD44 and mi R-145 were determined in gastric cancer cells. Quantitative real-time polymerase chain reaction was used to measure to the level of CD44 m RNA. A luciferase reporter assay and western blotting were performed to examine the effect of mi R-145 on CD44 expression. Tumor sphere and MTT assays were carried out to evaluate the self-renewal and chemo-resistance properties of gastric cancer cells.RESULTS The expression of CD44 was greatly increased and mi R-145 was decreased in gastric cancer cells that were highly enriched in cancer stem cells(CSCs). The results demonstrated that mi R-145 regulated CD44 by targeting directly the CD44 3'-untranslated region(3'-UTR). In gastric cancer cells, overexpression of mi R-145 repressed the activity of the CD44 3'-UTR, and disruption of mi R-145/CD44 3'-UTR interactions abrogated the silencing effects. In addition, mi R-145 inhibition stimulated CD44 3'-UTR activity and disruption of mi R-145/CD44 3'-UTR interactions abrogated this stimulatory effect. Enforced CD44 expression greatly increased tumor sphere formation and chemoresistance in gastric cancer cells. Furthermore, the inhibition of CSCs and the chemo-sensitivity of gastric cancer cells treated with mi R-145 were significantly abrogated by overexpression of CD44. CONCLUSION mi R-145 targeting of CD44 plays critical roles in the regulation of tumor growth and chemo-resistance in gastric cancer.

Relationship between expression of CD44v6 and nm23-H1 and tumor invasion and metastasis in hepatocellular carcinoma

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Lack of CD44 variant 6 expression in rectal cancer invasive front associates with early recurrence

AIM: To investigate the prognostic value of CD44 variant 6 (CD44v6), a membranous adhesion molecule, in rectal cancer. METHODS: Altogether, 210 rectal cancer samples from 214 patients treated with short-course radiotherapy (RT, n = 90), long-course (chemo) RT (n = 53) or surgery alone (n = 71) were studied with immunohistochemistry for CD44v6. The extent and intensity of membranous and cytoplasmic CD44v6 staining, and the intratumoral membranous staining pattern, were analyzed.RESULTS: Membranous CD44v6 expression was seen in 84% and cytoplasmic expression in 81% of the cases. In 59% of the tumors with membranous CD44v6 expression, the staining pattern in the invasive front was determined as "front-positive" and in 41% as "front-negative". The latter pattern was associated with narrower circumferential margin (P = 0.01), infiltrative growth pattern (P < 0.001), and shorter disease-free survival in univariate survival analysis (P = 0.022) when compared to the "front-positive" tumors. CONCLUSION: The lack of membranous CD44v6 in the rectal cancer invasive front could be used as a method to identify patients at increased risk for recurrent disease.

Expression of PCNA and CD44mRNA in colorectal cancer with venous invasion and its relationship to liver metastasis

AIM: To investigate the expression of proliferating cell nuclear antigen (PCNA) and CD44mRNA in colorectal cancer with venous invasion and its relationship with liver metastasis.METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of PCNA and CD44mRNA in 31 cases of colorectal cancer with venous invasion.RESULTS: Positive expression rates of PCNA and CD44mRNA in colorectal cancer were higher than those without liver metastasis (P＜0.05 and P＜0.01). In case of colorectal cancer with liver metastasis, strongly positive rates of PCNA and CD44mRNA were 94.1% and 70.6 %,respectively, significantly higher than those without liver metastasis. There was a positive relationship between the expressions of PCNA and CD44mRNA (r=0.67, P＜0.05).CONCLUSION: Detection of PCNA and CD44mRNA expression in colorectal cancer may be useful for evaluating liver metastasis of cancer cells.

Study on the Expression Levels of CXCR4, CXCL12, CD44, and CD147 and Their Potential Correlation with Invasive Behaviors of Pituitary Adenomas

Objective To evaluate the factors of CXCR4, CXCL12, CD44, and CD147 as early potential diagnostic biomarkers by determining their expression levels in invasive and non-invasive pituitary adenomas. Methods Fresh pituitary adenoma specimens were collected from 35 pituitary adenoma （21 invasive and 14 non-invasive） patients who underwent surgical treatment in our Neurosurgery Department between January and April of 2009. The expression levels of CXCR4, CXCL12, CD44, and CD147 were evaluated firstly by flow cytometry, fluorescence microscopy in single cell suspensions, and then by immunohistochemical staining of paraffin tissue sections. Results Flow cytometric analyses showed that the percentage of CXCR4- and CXCL12-positive cells from invasive pituitary adenomas （IPA） was significantly higher in the single cell suspensions than that from non-invasive pituitary adenomas （nlPA） （P〈O.05）. Immunohistochemical staining revealed that CXCR4 and CXCL12 staining index scores of the invasive pituitary adenomas were significantly higher than those of the non-invasive pituitary adenomas （P〈O.05）. In contrast, neither flow cytometry nor immunohistochemical staining demonstrated significant difference between CD44 and CD147 expression levels, respectively. Conclusion Expression levels of CXCR4 and CXCL12 are correlated with the invasiveness of pituitary adenomas. Therefore, rather than CD44 and CD147, CXCR4 and CXCL12 may potentially serve as biomarkers for early detection of pituitary adenomas.

Expression of proliferating cell nuclear antigen and CD44 variant exon 6 in primary tumors and corresponding lymph node metastases of colorectal carcinoma with Dukes'stage C or D

AIM: To study changes in characteristics of colorectal carcinoma during the metastatic process and to investigate the correlation between cell proliferation activity and metastatic ability of patients with Dukes′ stage C or D.METHODS: Formalin fixed and paraffin embedded materials of primary tumors and corresponding lymph node metastases resected from 56 patients with Dukes′ stage C or D of colorectal carcinoma were stained immunohistochemically with proliferating cell nuclear antigen (PCNA) and CD44variant exon 6 (CD44v6).RESULTS: Thirty-one of 56 patients (55.4 %) expressed PCNA in the primary sites and 36 of 56 patients (64.3 %)expressed PCNA in the metastatic lymph nodes. A significant relation in PCNA expression was observed between the primary site and the metastatic lymph node (0.010＜P＜0.025).Forty-one of 56 patients (73.2 %) expressed CD44v6 in the primary site and 39 of 56 patients (69.6 %) expressed CD44v6 in the metastatic lymph node. There was also an significant relationship of CD44v6 between the primary site and the metastatic lymph node (0.005＜P＜0.010). No difference was observed between expression of CD44v6 and PCNA in the primary site (0.250＜P＜0.500).CONCLUSION: This study partially demonstrates that tumor cells in metastatic lymph node of colorectal carcinoma still possess cell proliferation activity and metastatic ability of tumor cells in primary site. There may be no association between cell proliferation activity and metastatic ability in colorectal carcinoma.