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乳腺癌是女性常见恶性肿瘤和死亡原因之一,中国每年新发病例超过268.6万,占女性新发癌症的15.0%,发病率和死亡率均位居所有恶性肿瘤的第6位,严重威胁女性身心健康[1]。其中,三阴性乳腺癌(triple negative breast cancer,TNBC)是一种特殊类型的乳腺癌,约占所有乳腺癌的15%[2],是指雌激素受体(estrogen receptor,ER)、孕激素受体(progestogen receptor,PR)和人表皮生长因子受体2(human epidermal growth factor receptor-2,Her-2)均为阴性的乳腺癌[3]。TNBC发病年龄轻、侵袭性强、复发早、远处转移率高,其侵袭转移的分子机制研究成为关注重点。
缝隙连接蛋白(connexin,Cx)通过介导的细胞间通信广泛参与人体多种生物学事件的发生,诸如生物电活动的同步、组织分化发育、细胞生长及凋亡、代谢及营养物质传输等[4]。Cx的异常可导致包括肿瘤在内的多种疾病的发生[5-10]。人类乳腺组织中主要表达Cx26和Cx43[11],Cx26的下调或缺失一度被认为是乳腺癌发生进展过程中的重要分子事件[12-13],并且Cx26在淋巴结转移灶中的高表达被认为与预后不良密切相关[14-15]。Cx功能的发挥与多条信号通路有关[16-17],其中核转录因子κB(nuclear factor κB, NF-κB)是一种可与免疫球蛋白κ轻链基因增强子κB序列特异性结合的核蛋白因子,它在自身免疫性疾病、肿瘤(包括乳腺癌)等人类疾病的发生发展中发挥重要作用[18]。本文通过免疫组织化学方法探讨Cx26和NF-κB在TNBC原发灶和区域淋巴结转移灶中的表达,分析二者相关性及其与临床病理特征之间的关系。现作报道。
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Cx26蛋白主要分布于乳腺癌细胞的细胞膜,少量见于细胞质,多呈棕黄色颗粒分布;NF-κB主要定位于细胞核,多呈棕褐色染色(见图 1)。原发灶A组的Cx26整体阳性表达率为65.31%,其中A1组阳性表达率为40.91%,低于A2组的85.19%(P < 0.01);原发灶A组的NF-κB整体阳性表达率为53.06%,其中A1组阳性表达率为36.36%,低于A2组的66.67%(P < 0.05)(见表 1)。
分组 n Cx26 χ2 P NF-κB χ2 P 阳性 阴性 阳性 阴性 A1组 22 9 13 8 14 A2组 27 23 4 10.49 <0.01 18 9 4.47 <0.05 合计 49 32 17 26 23 表 1 Cx26和NF-κB在TNBC原发灶中的表达(n)
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淋巴结转移灶B组中Cx26和NF-κB的阳性表达率分别为81.48%(22/27)和77.78%(21/27)),与原发灶A组的65.31%和53.06%比较差异均无统计学意义(P>0.05)。但在亚组分析中发现,B组Cx26和NF-κB的阳性表达率均高于A1组中表达(40.91%和36.36%)(P < 0.01)(见表 2)。
分组 n Cx26 χ2 P NF-κB χ2 P 阳性 阴性 阳性 阴性 A1组 22 9 13 8 14 B组 27 22 5 8.59 <0.01 21 6 8.61 <0.01 合计 49 31 18 29 20 表 2 Cx26和NF-κB在TNBC原发灶A1组和淋巴结转移灶B组中的表达比较(n)
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Cx26蛋白的表达与TNBC病人的绝经状态、淋巴结是否转移有关(P < 0.01),表现为绝经前和有淋巴结转移者表达更高,而与病人的年龄、肿瘤大小、组织学分级和pTNM分期无关(P>0.05)。NF-κB蛋白的表达与pTNM分期和淋巴结是否转移有关(P < 0.01和P < 0.05),表现为分期越高和有淋巴结转移者表达更高,而与病人年龄、绝经状态、肿瘤大小和组织学分级无关(P>0.05)(见表 3)。
n Cx26 χ2 P NF-κB χ2 P - + - + 年龄/岁 <35 2 0 2 — >0.05* 1 1 — >0.05* ≥35 47 7 30 22 25 绝经状态 绝经前 27 5 22 6.95 <0.01 12 15 0.15 >0.05 绝经后 22 12 10 11 11 肿瘤大小/cm ≤5 45 16 29 0.02△ >0.05 21 24 0.16△ >0.05 >5 4 1 3 2 2 组织学分级/级 Ⅰ~Ⅱ 28 11 17 0.61 >0.05 12 16 0.44 >0.05 Ⅲ 21 6 15 11 10 pTNM分期/期 Ⅰ~Ⅱ 30 12 18 0.96 >0.05 19 11 8.35 <0.01 Ⅲ 19 5 14 4 15 淋巴结转移 阴性 22 13 9 10.49 <0.01 14 8 4.47 <0.05 阳性 27 4 23 9 18 *示Fisher′s确切概率法;△示校正χ2值 表 3 Cx26和NF-κB表达与TNBC临床病理特征的关系
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Cx26和NF-κB在TNBC原发灶中的表达无明显相关性(P>0.05),在淋巴结转移灶组织中两者表达呈明显正相关关系(r=0.663,P < 0.01)。
缝隙连接蛋白26和核转录因子κB在三阴性乳腺癌原发灶及淋巴结转移灶中的表达及临床意义
Expression levels of connexin 26 and NF-κB in primary lesion and metastatic lymph nodes of triple negative breast cancer, and its clinical significance
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摘要:
目的探讨缝隙连接蛋白26(connexin 26,Cx26)和核转录因子κB(nuclear factor κB,NF-κB)在三阴性乳腺癌(triple negative breast cancer,TNBC)病人原发灶和淋巴结转移灶中的表达及临床意义。 方法回顾性收集经术后病理和分子分型确诊为TNBC的存档石蜡组织,原发灶共49例,其中无区域淋巴结转移者22例(A1组),有区域淋巴结转移者27例(A2组);将27例淋巴结转移灶作为B组。应用免疫组织化学SP染色法,检测各组中Cx26和NF-κB的表达情况,并分析二者表达的相互关系及其与临床病理特征的相关性。 结果原发灶中A2组Cx26和NF-κB阳性表达率均高于A1组(P < 0.01和P < 0.05);转移灶B组Cx26和NF-κB阳性表达率均高于原发灶A1组(P < 0.01)。Cx26蛋白表达与TNBC病人绝经状态、淋巴结转移有关(P < 0.01),NF-κB表达与pTNM分期、淋巴结转移有关(P < 0.01和P < 0.05)。在淋巴结转移灶中,Cx26与NF-κB表达呈明显正相关关系(r=0.663,P < 0.01)。 结论Cx26和NF-κB共同参与TNBC的浸润和转移,可作为预测TNBC转移复发及评估临床预后的有效指标。 Abstract:ObjectiveTo investigate the expression levels of connexin 26(Cx26) and nuckar factor κB(NF-κB) in primary lesion and metastatic lymph nodes of triple negative breast cancer(TNBC), and its clinical significance. MethodsThe TNBC paraffin tissues diagnosed by postoperative pathology and molecular classification were collected.Forty-nine cases of primary lesions and 27 cases of lymph nodes metastatic lesions were divided into group A and group B, respectively.Group A included 22 cases without regional lymph node metastasis(group A1) and 27 cases with regional lymph node metastasis(group A2).The expression levels of Cx26 and NF-κB in all cases were examined using immunohistochemical staining.The relationship between Cx26 and NF-κB, and their correlations with clinical features were analyzed. ResultsThe positive expression rates of Cx26 and NF-κB in group A2 were higher than those in group A1(P < 0.01 and P < 0.05).The positive expression rates of Cx26 and NF-κB in group B were higher those in group A1(P < 0.01).The expression level of Cx26 was correlated with the menopause state and lymph node metastasis(P < 0.01), and the NF-κB level was correlated with pTNM staging and lymph node metastasis(P < 0.01 and P < 0.05).In lymph nodes metastatic lesions, the expression of Cx26 was positively correlated with the expression of NF-κB(P < 0.01). ConclusionsCx26 and NF-κB may co-participate in the invasion and metastasis of TNBC, and can be used as an effective indicator to predict TNBC metastasis and recurrence and evaluate clinical prognosis. -
Key words:
- triple negative breast cancer /
- tumor metastasis /
- connexin 26 /
- nuclear factor-κB
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表 1 Cx26和NF-κB在TNBC原发灶中的表达(n)
分组 n Cx26 χ2 P NF-κB χ2 P 阳性 阴性 阳性 阴性 A1组 22 9 13 8 14 A2组 27 23 4 10.49 <0.01 18 9 4.47 <0.05 合计 49 32 17 26 23 表 2 Cx26和NF-κB在TNBC原发灶A1组和淋巴结转移灶B组中的表达比较(n)
分组 n Cx26 χ2 P NF-κB χ2 P 阳性 阴性 阳性 阴性 A1组 22 9 13 8 14 B组 27 22 5 8.59 <0.01 21 6 8.61 <0.01 合计 49 31 18 29 20 表 3 Cx26和NF-κB表达与TNBC临床病理特征的关系
n Cx26 χ2 P NF-κB χ2 P - + - + 年龄/岁 <35 2 0 2 — >0.05* 1 1 — >0.05* ≥35 47 7 30 22 25 绝经状态 绝经前 27 5 22 6.95 <0.01 12 15 0.15 >0.05 绝经后 22 12 10 11 11 肿瘤大小/cm ≤5 45 16 29 0.02△ >0.05 21 24 0.16△ >0.05 >5 4 1 3 2 2 组织学分级/级 Ⅰ~Ⅱ 28 11 17 0.61 >0.05 12 16 0.44 >0.05 Ⅲ 21 6 15 11 10 pTNM分期/期 Ⅰ~Ⅱ 30 12 18 0.96 >0.05 19 11 8.35 <0.01 Ⅲ 19 5 14 4 15 淋巴结转移 阴性 22 13 9 10.49 <0.01 14 8 4.47 <0.05 阳性 27 4 23 9 18 *示Fisher′s确切概率法;△示校正χ2值 -
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