CN116298292B - Gng2蛋白的检测试剂在制备诊断和预后结肠癌的产品中的用途 - Google Patents
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Abstract
本发明属于医学诊断技术领域，具体涉及GNG2蛋白的检测试剂在制备诊断和预后结肠癌的产品中的用途。所述GNG2蛋白的NCBI登录号为NP_001230702，所述诊断和预后的受试者为人，所述检测的样本为受试者的组织。本发明所述的GNG2蛋白高表达与结肠癌的发生及恶性生长行为相关，相较于健康对照，GNG2蛋白高表达指示受试者患有结肠癌或有患结肠癌的风险；结肠癌患者组织样本中GNG2蛋白表达升高，判定为不良预后；结肠癌患者组织样本中GNG2蛋白表达降低，判定为良好预后。结果提示GNG2蛋白是一个潜在的结肠癌治疗靶点。
Description
技术领域
本发明属于医学诊断技术领域，具体涉及GNG2蛋白的检测试剂在制备诊断和预后结肠癌的产品中的用途。
背景技术
结直肠癌是恶性程度非常高的肿瘤之一，其发病率和死亡率有明显增高的趋势。全球每年约有120万的新发病例，约有50万人死于结直肠癌，已成为第三大类癌症死因。随着居民生活水平的不断提高、饮食习惯的改变，结直肠癌的发病率呈逐年上升趋势。当前，结直肠癌发病率仅次于肺癌、胃癌，位于第3位，是严重威胁人类健康的疾病，外科手术是直肠癌根治首选的治疗方法。CEA、CA199等传统肿瘤标志物敏感性较差，在早期结肠癌患者中阳性率低，不利于早期筛查。目前，结肠癌的诊断主要依赖肠镜及腹部CT等方法，大多数患者发现时已处于肿瘤晚期，失去了最佳的治疗时机，因此寻找特异的关键分子及治疗靶点对结肠癌的早期诊断、预后判断、新药开发和临床治疗具有重要意义。
GNG2(G protein gamma 2subunit)是异源三聚体G蛋白中的一个γ2小亚基,NCBI登录号为NP_001230702。研究报道，异源三聚体G蛋白参与多种细胞信号的转导，在细胞增殖、分化和血管生成中发挥重要作用，为多种疾病的化疗提供了潜在的分子靶点。GNG2作为异三聚体G蛋白的一部分，已有报道指出其能通过独立地抑制AKT和FAK的活性进而抑制人类恶性黑色素瘤细胞的增殖和侵袭。此外，大数据分析显示，GNG2还在IgA肾病、唐氏综合征、嘌呤代谢等多种疾病和生物学过程中发挥关键作用。尽管如此，GNG2蛋白在结肠癌中的表达情况及其临床意义目前尚不清楚。
发明内容
本发明的目的在于寻求有效的预后监测指标，以期为提高结肠癌患者的预后奠定基础。
为实现上述目的，本发明提供如下技术方案：
GNG2蛋白的检测试剂在制备诊断和预后结肠癌的产品中的用途，所述GNG2蛋白的NCBI登录号为NP_001230702。
进一步的，上述用途中，所述诊断和预后的受试者为人。
上述用途中，作为一种优选的实施方案，所述检测的样本为受试者的组织。
进一步的，相较于健康对照，GNG2蛋白高表达指示受试者患有结肠癌或有患结肠癌的风险。
进一步的，结肠癌患者组织样本中GNG2蛋白表达升高，判定为不良预后；结肠癌患者组织样本中GNG2蛋白表达降低，判定为良好预后。
上述用途中，所述检测试剂为定量检测所述GNG2蛋白表达量的试剂。
进一步的，上述用途中，所述检测的方法为放射方法、免疫方法、荧光方法、流式荧光法、胶乳比浊法、生化法、酶法、杂交法、气质联用法、液质联用法、层析法、化学发光方法、磁电法或光电转换方法。
与现有技术相比，本发明具有如下有益效果：
本发明研究结果表明：GNG2蛋白在结肠癌组织中的表达明显高于癌旁组织(p<0.001)；GNG2蛋白表达与病理分级、PDL-1蛋白密切相关(p＜0.05)；癌组织GNG2蛋白高表达与结肠癌的不良预后显著正相关(p＝0.02459)。该结果提示GNG2蛋白高表达与结肠癌的发生及恶性生长行为相关，可作为预后不良的判断指标，且提示GNG2蛋白是一个潜在的结肠癌治疗靶点。
附图说明
图1为GNG2在结肠癌及癌旁组织中的表达比较。***Statistically significant(p<0.001)。
图2为GNG2表达量与结肠癌患者预后的相关性。
具体实施方式
下面结合附图和具体实施例对本发明进行详细说明，但不应理解为本发明的限制。如未特殊说明，下述实施例中所用的技术手段为本领域技术人员所熟知的常规手段，下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1
1材料与方法
1.1组织样本来源
104例结肠癌患者样本来源于上海芯超生物科技有限公司的组织样本库，手术时间为2006年7月至2007年5月，最终随访时间均为2015年7月。所有病例均病理确诊为结肠癌，并且未接受任何术前治疗。男性58例，女性45例，1例未知。年龄24-90岁，中位年龄57岁。每例均包含癌组织和相匹配的距离癌1.5cm的癌旁组织，104例结直肠癌患者的临床病理资料见表1。
表1 104例结直肠癌患者的临床病理资料
1.2组织芯片制作
由上海芯超生物科技有限公司完成组织芯片的制作。所有供体组织蜡块行常规病理切片后进行HE染色，由病理专家作二次诊断，并在HE切片上标记典型病理部位。使用组织芯片制作仪(BeecherInstruments.Inc)在受体蜡块(空白蜡块)上打孔(直径1.5mm)，然后根据HE片上的标记范围在供体组织蜡块相应位置获取目标组织芯放入受体蜡块阵列孔中，重复上述步骤，最终制作完成1个结肠癌组织和癌旁组织匹配的180点阵列块(HCol-Ade180Sur-06)。采用切片机(Leica，德国)以4-5um厚度连续切片，切片裱附在经过防脱片处理的进口载玻片上制成组织芯片。
1.3免疫组织化学实验
EnVision二步法进行免疫组织化学实验。在光学显微镜下随机观察3个高倍镜视野，记不少于3×100个细胞中的阳性细胞数，计算阳性细胞数占整个细胞数的染色阳性率。以“染色强度评分”和“染色阳性率评分”的乘积为总评分进行分组，＜12分为抗体低表达组，≥12分为抗体高表达组。
1.4统计学分析
GNG2蛋白在结肠癌和癌旁组织中的表达情况采用卡方检验分析。GNG2表达与结肠癌患者临床指标相关性采用卡方检验分析。GNG2蛋白与结肠癌预后的相关性分析：采用Kaplan-Meier生存分析法和log-rank统计检验进行生存期单因素分析。p<0.05有统计学意义。
2实验结果
2.1GNG2蛋白在结肠癌和癌旁组织中的表达分析
免疫组化实验结果显示：GNG2蛋白在结肠癌和癌旁组织的细胞浆表达。卡方检验分析显示：GNG2在结肠癌中表达显著高于癌旁组织(p<0.001)，分析结果见图1。
2.2GNG2表达与结肠癌患者临床指标相关性
卡方检验分析显示：GNG2表达与病理分级、PDL-1密切相关(p＜0.05)。此外，GNG2表达与年龄、性别、T分期、N分期、TNM分期、MLH1、MSH6、MSH2、PMS2之间无明显统计学关系(P＞0.05)，见表2。
表2GNG2表达与结肠癌患者临床指标相关性
2.3TRIM蛋白表达与结肠癌预后的相关性分析
结肠癌病人的随访情况如下：手术时间为2006年7月至2007年5月，最终随访时间为2015年7月。随访期间，有63例病人死于结肠癌，中位随访时间28个月(1到87个月)；有41例病人仍存活，中位随访时间101个月(98到108个月)。采用Kaplan-Meier生存分析法和log-rank统计检验进行生存期单因素分析显示：癌组织GNG2表达低的结肠癌病人拥有更长的总生存期(p＝0.02459)，其5年生存率显著高于GNG2表达高的结肠癌病人(图2)。
尽管已描述了本发明的优选实施例，但本领域内的技术人员一旦得知了基本创造性概念，则可对这些实施例作出另外的变更和修改。所以，所附权利要求意欲解释为包括优选实施例以及落入本发明范围的所有变更和修改。
显然，本领域的技术人员可以对本发明进行各种改动和变型而不脱离本发明的精神和范围。这样，倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内，则本发明也意图包含这些改动和变型在内。
Claims (3)
1.GNG2蛋白的检测试剂在制备预后结肠癌的产品中的用途，其特征在于，所述GNG2蛋白的NCBI登录号为NP_001230702，所述预后的受试者为人，所述检测的样本为人的组织。
2.根据权利要求1所述的用途，其特征在于，结肠癌患者组织样本中GNG2蛋白表达升高，判定为不良预后；结肠癌患者组织样本中GNG2蛋白表达降低，判定为良好预后。
3.根据权利要求1所述的用途，其特征在于，所述检测试剂为定量检测所述GNG2蛋白表达量的试剂。
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