DETECTION OF SBEM-MRNA IN PERIPHERAL BLOOD OF PATIENTS WITH BREAST CANCER AND ITS CLINICAL SIGNIFICANCE

Objective： The aim of the present study is to explore the expression of a specific marker of breast cancer, small breast epithefial mucin （SBEM） mRNA, in peripheral blood and to investigate its significance. Methods： The expressions of SBEM-mRNA in peripheral blood of 67 patients with breast cancer, 16 patients with benign breast disease, and 20 normal healthy volunteers were detected with nested reverse transcription-polymerase chain reaction （Nested-RT-PCR）. Results： SBEMomRNA was negative in healthy individuals and patients with benign breast tumor, but 50.7%（34/67） of breast cancer patients showed positive expression of SBEM-mRNA in peripheral blood, of whom 25%（2/8） were in stage Ⅰ, 45.8%（11/24） in stage Ⅱ, 43.75%（11/24） in stage Ⅲ and 73.7（14/19） in stage Ⅳ. The positive rate in stage Ⅳ was higher than that in stage Ⅰ, Ⅱ, Ⅲ （P〈0.05）. Expressions of SBEM-mRNA had no correlation with age, carcinoma size, pathological type, ER and PR state （P〉0.05）. Concision： SBEM-mRNA is specifically expressed in breast cancer and it may act as a marker for the detection of of breast cancer.

Expression of CK19 mRNA and MUC-1 mRNA in the peripheral blood of patients with colorectal cancer and their clinical significances

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.

Quantitative detection of peripheral MDR genes in breast cancer before chemotherapy and their clinical significances

Objective： To explore expressions of multidrug resistance gene （MDR） product P170 and peripheral blood MDR1 mRNA in breast cancer tissue and their clinical significances. Methods： Fluorescent quantitative RT-PCR and immunohistochemistry were used to detect peripheral blood MDR1 mRNA before chemotherapy in breast cancer tissue samples from 32 cases with P170 positive （trial group） and 11 cases with P170 negative （control group）. Results： There were 14 cases （43.75%） peripheral blood MDR1 mRNA positive and 18 cases （56.25%） negative in 32 cases P170 positive breast cancer patients, while there were 6 cases （54.55%） peripheral blood MDR1 mRNA positive and 5 cases （45.45%） negative in 11 cases P170 negative breast cancer patients; there wasn＇t a significance difference with peripheral blood MDR1 mRNA express rate in P170 positive （43.75%） group and P170 negative （54.55%） group （x2 = 0.383, P 〉 0.05）. There were 12 cases （37.50%） Her-2 positive and 20 cases （62.50%） negative in P170 positive group; there were 2 cases （18.18%） Her-2 positive and 9 cases （81.82%） negative in P170 negative group; there wasn＇t a significance difference with Her-2 positive rate in P170 positive group （37.50%） and P170 negative （18.18%） group （χ^2= 1.391, P 〉 0.05）. There were 13 cases （40.63%） ER positive and 19 cases （59.37%） negative in P170 positive group; there were 7 cases （63.64%） ER positive and 4 cases （36.36%） negative in P170 negative group; there wasn＇t a significance difference with ER positive rate in P170 positive group （40.63%） and P170 negative （63.64） group （χ^2 = 1.742, P 〉 0.05）. There were 20 cases （62.50%） PR positive and 12 cases （37.50%） negative in P170 positive group; there were 4 cases （36.36%） PR positive and 7 cases （63.64%） negative in P170 negative group; there wasn＇t a significance difference with PR positive rate in P170 positive group （62.50%） and P170 negative （36.36%） group （χ^2 = 2.267, P 〉 0.05）. There were 9 cases （28.12%） with lymphaden metastasis and 23 cases （71.88%） without lymphaden metastasis in P170 positive group; there were 5 cases （45.45%） with lymphaden metastasis and 6 cases （54.55%） without lymphaden metastasis in P170 negative group; there wasn＇t a significance difference with lymphaden metastasis rate in P170 positive group （28.12%） and P170 negative （45.45%） group （χ^2 = 1.120, P 〉 0.05）. Conclusion： 1） There were possibly MDR expressing infrequently in carcinoma tissue and peripheral blood before chemotherapy. 2） FQ-RT-PCR for detecting of peripheral blood MDR1 mRNA is special, sensitive and reliable. It can be used as new molecular biology diagnostic maker dynamic detecting peripheral blood MDR1 mRNA for regulating chemotherapy proposal and elevating chemotherapy effect.

肺癌患者外周血循环细胞CEA-mRNA RT-PCR检测的意义

目的 了解肺癌患者外周血CEA -mRNA的水平。方法 采用实时荧光RT- PCR技术,检测肺癌患者78例,肺部良性疾病(BLD)患者70例,健康志愿者30例外周血CEA- mRNA的水平。结果 78例肺癌患者外周血CEA- mRNA的平均水平为(10 2 38. 4±185 2 6 . 3)copies/ml,其中4 3例阳性、阳性率为5 5 . 1%。BLD患者外周血CEA mRNA平均水平为(82. 7 4±6 0. 3 7)copies/ml,其中阳性2例,阳性率为6 . 7%。健康对照未检测到CEA -mRNA。CEA -mRNA水平与肿瘤病理分期和分化程度相关;CEA -mRNA阳性组患者血清CEA蛋白的阳性率与CEA mRNA阴性组相比差异无显著性(P>0 0 5 )。结论 外周血CEA mRNA可作为肺癌辅助诊断指标,定量检测外周血CEA- mRNA可作为肺癌微转移的一个监测指标。

SBEM-mRNA与CEA在乳腺癌患者外周血表达的临床意义

目的:通过检测人类原发性乳腺癌患者外周血中乳腺小粘蛋白(SBEM)mRNA和癌胚抗原(CEA)的表达情况,同时结合患者其他相关临床病理学资料,探讨二者检测在乳腺癌患者中的临床意义。方法:采用逆转录-聚合酶反应(RT-PCR)和酶联免疫反应(ELESA)方法分别检测60例乳腺癌患者外周血中SBEM-mRNA和CEA的表达情况,另抽取20例乳腺纤维腺瘤患者和10例健康人的外周血标本加以检测,作为对照研究组。采用SPSS10.0软件进行统计学处理。结果:在60例乳腺癌患者外周血中SBEM-mRNA和CEA的阳性表达率分别为53.3%和31.7%。而在对照组检测中,CEA在纤维腺瘤患者外周血中阳性率为5.0%,在健康人的外周血中为阴性表达;SBEM-mRNA在二者中均为阴性表达。统计学分析发现,乳腺癌患者中SBEM-mRNA和CEA的阳性表达均与腋淋巴结转移情况、肿瘤TNM分期差异有显著性意义(P<0.05),而与患者的其他临床病理学特征的差异无统计学意义(P>0.05)。对比研究表明,二者联合检测比单项检测有更高的敏感度和准确性(P<0.05)。结论:乳腺癌患者外周血中SBEM-mRNA和CEA的检测对指导乳腺癌患者的治疗和判断预后具有重要意义,二者联合检测在提高对乳腺癌微转移敏感度和准确性的同时,又保持了较高的特异性,是较为理想的检测组合方式。

RT-PCR检测乳腺癌患者外周血HMGA2mRNA表达的意义

目的探讨乳腺癌患者外周血中HMGA2mRNA的表达,建立检测外周血早期诊断乳腺癌微转移的方法。方法采取72例乳腺癌和41例乳腺良性疾病患者的外周血,运用RT-PCR技术进行mRNA的检测。结果72例乳腺癌患者HMGA2mRNA有27例表达,总阳性率为37.50%。41例乳腺良性疾病者HMGA2mRNA无1例表达。两组有统计学意义(χ2=7.35,P<0.001)。结论HMGA2可以用于临床检测乳腺癌患者外周血中微转移,有可能成为判断预后和指导治疗的指标。

乳腺癌患者外周血CEA mRNA的表达

目的 检测乳腺癌患者外周血CEAmRNA的表达 ,建立取材于外周血早期诊断乳腺癌转移的方法。方法 用巢式RT -PCR检测乳腺癌患者和乳腺良性肿瘤患者的外周血中CEAmRNA的表达。结果 5 6例乳腺癌患者CEAmRNA有 2 1例表达 ,阳性率为 3 7.5 0 % ;42例乳腺良性疾病者CEAmRNA没有一例表达。两组有高度显著性差异 (χ2 =2 0 .0 4 ,P <0 .0 1 ) 结论 巢式RT -PCR检测CEAmRNA阳性结果提示其具有形成转移的倾向 。

非小细胞肺癌患者外周血CEA mRNA表达与临床及病理的相关性研究

目的分析外周血CEAmRNA表达与非小细胞肺癌(NSCLC)的临床病理相关性,探讨CEAmRNA定量检测对非小细胞肺癌治疗及预后的临床意义。方法以CEAmRNA为靶基因,运用逆转录-实时定量PCR技术,定量检测49例非小细胞肺癌患者治疗前后、10例良性肺部病变患者、10例健康人员外周血CEAmRNA的表达情况。结果①10例健康人员及10例肺部良性疾病患者外周血中CEAmRNA均无表达,NSCLC患者外周血中CEAmRNA表达阳性率77.6%,5例NSCLC组织均检测到CEAmRNA。②Ⅰ、Ⅱ、Ⅲ、Ⅳ期NSCLC患者外周血中CEAmRNA阳性检出率分别为42.9%(3/7)、71.4%(5/7)、83.3%(15/18)、88.2%(15/17),随着病程进展,表达阳性率增高,Ⅰ、Ⅱ期与Ⅲ、Ⅳ期比较差异有显著性(χ2=4.689,P<0.05;t=-2.296,P=0.026)。③NSCLC患者手术前后外周血中CEAmRNA变化差异显著(t=2.267,P=0.033)。④Ⅲb期及Ⅳ期NSCLC患者放、化疗前后外周血CEAmRNA变化差异显著(t=2.584,P=0.016),临床疗效不同的患者其外周血中CEAmRNA表达量变化不同(F=7.228,P=0.004;γ=0.629,P=0.01)。结论逆转录-实时定量PCR法能特异、灵敏地检测非小细胞肺癌患者外周血中CEAmRNA表达。非小细胞肺癌早期可能就有隐匿性的远处转移,检测外周血中癌细胞可能对判断疗效及改进治疗方案具有积极地临床意义。

乳腺癌外周血细胞角蛋白19 mRNA的检测及临床价值

目的应用荧光定量RT-PCR检测乳腺癌外周血中的表达情况,并评价其表达的临床意义。方法用荧光定量RT-PCR方法检测49例乳腺癌患者外周血CK19mRNA表达。结果49例乳腺癌患者外周血中CK19mRNA检测阳性28例,阳性率57.1%,统计结果显示乳腺癌患者外周血CK19mRNA的表达阳性率在患者不同年龄、病理类别、临床分期、ER、PR状态的各组差异无显著性(P>0.05),在淋巴结转移状态间有显著差异。结论CK19mRNA可作为RT-PCR法检测乳腺癌患者外周血微转移的分子标志物,有助于早期预测乳腺癌发生血道微转移,指导临床治疗。

hMAM mRNA表达情况与乳腺癌的关系

目的探讨人乳腺珠蛋白(h MAM)m RNA在乳腺癌患者外周血中的表达情况及意义。方法选取87例乳腺癌患者、70例乳腺良性肿瘤患者和50例健康志愿者,采用巢式-逆转录-聚合酶链反应检测其外周血h MAM m RNA的表达水平。结果乳腺癌患者的外周血h MAM m RNA阳性率为54.02%,高于乳腺良性肿瘤患者的1.43%和健康志愿者的0,差异均有统计学意义(P﹤0.05);乳腺良性肿瘤患者和健康志愿者的外周血h MAM m RNA阳性率比较,差异无统计学意义(P﹥0.05)。Ⅲ~Ⅳ期、有淋巴结转移的乳腺癌患者的h MAM m RNA阳性率高于Ⅰ~Ⅱ期和无淋巴结转移的患者(P﹤0.05)。乳腺癌患者化疗后的外周血h MAM m RNA阳性率为18.39%,低于化疗前的54.02%(P﹤0.05)。结论乳腺癌患者外周血h MAM m RNA的阳性表达与TNM分期和淋巴结转移情况有关,可为临床诊断和预后判断提供依据。

大肠癌患者外周血中细胞角蛋白19和癌胚抗原的表达及临床意义

目的:应用巢式逆转录聚合酶链反应(nested RT-PCR)技术检测大肠癌患者外周血中CK19 mRNA、CEA mRNA的表达及临床意义,探讨其作为大肠癌微转移检测分子标记物的可能性。方法:应用巢式RT-PCR技术检测20例健康人、20例大肠腺瘤疾病患者和90例大肠癌患者外周血CK19 mRNA和CEA mRNA的表达。结果:大肠癌组CK19 mRNA和CEA mRNA表达的阳性率为58.89%(53/90)和53.33%(48/90);20例健康人外周血中无CK19 mRNA和CEA mRNA的表达;20例大肠腺瘤疾病患者有1例CK19 mRNA的表达,阳性率为5.00%(1/20),无CEA mRNA的表达。大肠癌患者中CK19 mRNA、CEA mRNA的阳性率与肿瘤分期及肿瘤转移有显著相关性(P<0.05)。结论:CK19 mRNA和CEA mRNA作为检测大肠癌患者外周血微转移的分子标记物具有良好的敏感度和特异性,同时可作为判断大肠癌患者的预后指标。