乳腺癌 C:C—erbB—2、p—Akt蛋白在乳腺癌中的表达水平及其在保存NAC的乳腺癌手术中的临床意义研讨

来源:中国现代医生 ·2018年12月29日 11:26 浏览量:0

盛贤达 郭宇 鲁杨 王金秋

[摘要] 意图 探討C-erbB-2、p-Akt蛋白在乳腺癌中的表达水平及其在保存NAC乳腺癌手术中的临床含义。 办法 自2010年6月~2011年3月,随机挑选86例经穿刺病理确诊确诊乳腺非滋润性导管癌,且契合保存NAC乳腺癌手术的女人患者为患者组。在我院健康体检中心随机挑选86例健康女人为健康对照组。对患者组患者进行保存NAC乳腺癌手术,术后对癌安排及癌旁正常安排进行免疫组化染色。检测C-erbB-2、p-Akt蛋白在癌安排及癌旁正常安排中的表达水平。运用ELISA法对患者手术前后及健康对照组外周血的C-erbB-2、p-Akt表达水平进行检测,记载各组数据并进行统计学剖析。依照C-erbB-2、p-Akt表达状况分为双阳性、双阴性以及单阳性。运用Kaplan-Meier法剖析患者生计预后。 成果 在乳腺癌安排及癌旁正常安排中,C-erbB-2、p-Akt蛋白均有表达,但在癌安排表达水平显着高于正常安排,差异具有统计学含义(P<0.05)。乳腺癌患者外周血C-erbB-2、p-Akt与健康对照组比较均显着升高。Kaplan-Meier法剖析患者生计预后发现,C-erbB-2、p-Akt均阳性表达者生计率显着下降,总生计期显着缩短,差异具有统计学含义(P<0.05)。 定论 乳腺癌患者外周血C-erbB-2、p-Akt水平显着高于健康对照组,C-erbB-2、p-Akt对患者承受保存NAC乳腺癌术后生计期有重要的临床含义。

[关键词] 乳腺癌;C-erbB-2;p-Akt;乳头乳晕复合体

[中图分类号] R737.9 [文献标识码] A [文章编号] 1673-9701(2017)12-0001-04

[Abstract] Objective To investigate the expression level of C-erbB-2 and p-Akt proteins in breast cancer and its clinical significance in breast cancer surgery reserving NAC. Methods From June 2010 to March 2011, 86 patients who were diagnosed with noninvasive ductal carcinoma of the mammary gland by biopsy pathological diagnosis and eligible for breast cancer surgery reserving NAC were randomly selected as the patients group. Another 86 healthy women were selected as the control group, randomly selected from the Physical Examination Center in our hospital. The enrolled patients were given the breast cancer surgery reserving NAC. Immunohistochemical staining was performed on cancer tissues and adjacent normal tissues after the surgery. The expression levels of C-erbB-2 and p-Akt in cancer tissues and adjacent normal tissues were detected, and the expression levels of C-erbB-2 and p-Akt in the peripheral blood before and after the surgery and in the healthy control group were detected by ELISA. The data of each group were recorded and analyzed statistically. According to the expression of C-erbB-2 and p-Akt, the expression was divided into double positive, double negative and single positive. Kaplan-Meier method was used to analyze the prognosis of patients. Results C-erbB-2 and p-Akt proteins were expressed in breast cancer tissues and adjacent normal tissues, but the expression levels in cancer tissues were significantly higher than those in normal tissues. The difference was statistically significant(P<0.05). The levels of C-erbB-2 and p-Akt in the peripheral blood of patients with breast cancer were significantly higher than those in the healthy control group. Kaplan-Meier method was used to analyze the survival prognosis, indicating that the survival rate of the patients with positive C-erbB-2 and p-Akt was significantly decreased, the overall survival time was significantly shorter, and the difference was statistically significant (P<0.05). Conclusion The levels of C-erbB-2 and p-Akt in peripheral blood of the patients with breast cancer are significantly higher than those in healthy control group, and C-erbB-2 and p-Akt have important clinical significance in the postoperative survival period of the patients receiving breast cancer reserving NAC.

[Key words] Breast cancer; C-erbB-2; p-Akt; Nipple-areola complex (NAC)

乳腺癌是指乳腺上皮细胞的恶性肿瘤,占女人一切恶性肿瘤的20%以上,成为要挟女人身体健康的重要杀手[1]。跟着乳腺癌患者对日子质量的要求越来越高,患者对术后漂亮程度的要求也越来越高,保存乳头乳晕复合体的乳腺癌彻底治愈术也得到了广泛的运用[2]。保存乳头乳晕复合体(nipple-areolar complex, NAC)对乳腺癌患者乳房重建的美容效果和术后日子质量有重要含义。可是保存NAC的乳腺癌术后危险即复发研讨尚不清晰[3]。因而,评论保存NAC的乳腺癌术后标志物关于研讨其发病机制以及疾病预后具有重要的临床含义[4]。本研讨首要运用C-erbB-2、p-Akt蛋白聯合点评保存NAC手术后患者的生计预后,并进一步评论C-erbB-2、p-Akt蛋白的临床含义。

1 材料与办法

1.1临床材料

自2010年6月~2011年3月,随机挑选86例经穿刺病理确诊确诊乳腺非滋润性导管癌,且契合保存NAC乳腺癌手术的女人患者为患者组。在我院健康体检中心随机挑选86例健康女人为健康对照组。两组年纪、BMI等一般材料比较,差异无统计学含义(P>0.05),具有可比性。见表1。

1.2乳腺癌患者入组规范[5]

(1)自2010年6月~2011年3月,在我院经过病理学确诊确诊乳腺癌的患者。(2)入组患者均为女人,年纪42~67岁,临床材料及病理材料均完好;(3)入组患者均需求契合保存乳头乳晕复合体乳腺癌手术规范,即:①适用于Ⅰ、Ⅱa期乳腺癌患者;②非中心区乳腺癌,肿瘤边际距乳晕边际直线间隔≥3 cm;③肿瘤最大径≤3 cm;④乳头无溢液、无湿疹样改动、无内陷及倾斜等反常体现;⑤乳晕及边际皮肤无桔皮样改动;⑥保存的乳头乳晕区皮下安排取样冰冻病理陈述无癌细胞滋润。(4)患者术前未承受放疗或化疗等医治。

1.3 乳腺癌患者扫除规范

(1)兼并其他体系的恶性肿瘤,不包含乳腺癌搬运到其他病灶者;(2)不能清晰确诊者;(3)认知妨碍或患精神疾病者;(4)某些原因导致未能获得病理样本者;(5)患者或家族不合作;(6)某些原因导致半途退出本研讨者;(7)患者一般状况较差,不合适进行检查及医治者。(8)患者术前承受过放疗或化疗等医治。

1.4免疫组化办法

乳腺癌安排白腊切片脱蜡至水,3%H2O2,20℃孵育10 min,蒸馏水冲刷,PBS浸泡5 min重复两次;运用5%PBS稀释的山羊血清(Life Technologies,美国)关闭,20℃孵育10 min,倒去山羊血清,滴加一抗(anti-C-erb-2,anti-p-Akt,ab60709,Abcam,美国),在37℃下孵育1~2 h或在4℃冰箱中过夜保存;将“生物素符号二抗”(Perkin-Elmer)工作液滴加适量到样品上,37℃孵育10 min;滴加辣根过氧化物酶(Sigma,S7571,美国),37℃孵育10 min;向1 mL DAB Plus Substrate(TA-125-HDX,Thermo Scientific,美国)中滴加1~2滴DAB Plus Chromogen(TL-125-HD,Thermo Scientific,美国),混匀后滴加到切片上,37℃孵育10 min后,冲刷封片。

1.5 ELISA检测办法

选用酶联免疫吸附(ELISA)试验检测患者血清C-erbB-2、p-Akt蛋白的表达水平,本研讨一切试剂均购于武汉博士德生物工程有限公司,操作彻底依照说明书进行。

1.6 手术办法

局麻下小切断(4~5 cm)切除肿瘤,病理成果陈述为癌后,在全麻下行保存NAC的乳腺癌改进彻底治愈术。以活检切断为轴,在距切断皮肤3~5 cm左右作一梭形切断,切除乳腺肿瘤外表的部分皮肤,向四周游离皮瓣,皮瓣下留有薄层脂肪,保存乳头乳晕,其基底部留有薄层腺体安排,术中取样冰冻切片。由内下沿胸大肌外表向上外连同胸肌筋膜同时切除至胸大肌外侧缘, 依据铲除腋淋巴结的需求而延伸切断或另行挑选切断,拉起胸大肌外侧缘,铲除腋窝淋巴结,整块切除手术标本。如确诊为滋润性癌者,术后需求承受化疗。

1.7 调查目标

调查患者手术前后ELISA检测、免疫组化检测成果及术后5年生计率。

1.8 统计学办法

对所搜集材料选用SPSS 15.0统计学软件处理,计量材料用(x±s)表明,组间比照选用独立样本t查验;计数数据用例数或构成比表明,选用χ2查验。选用Log Rank时序查验以及COX多要素剖析对患者临床预后进行剖析。P<0.05为差异具有统计学含义。

2 成果

2.1 患者组及健康对照组一般临床材料比较

患者组及健康对照组一般临床材料比较,两组无显着统计学差异(P>0.05)。见表1。乳腺癌患者病理类型均为导管癌。

2.2 ELISA检测成果

手术前后运用ELISA法检测患者外周血C-erbB-2、p-Akt蛋白表达水平发现,术前患者组外周血C-erbB-2、p-Akt蛋白水均匀显着高于健康对照组,差异具有统计学含义(P<0.05)。术后患者组外周血C-erbB-2蛋白水平显着减低至(1.805±0.001)ng/mL,低于术前(2.814±0.825)ng/mL(P<0.05),但与健康对照组比较,水平显着较高,差异具有统计学含义(P<0.05)。p-Akt成果发现,术后患者组血清p-Akt水平显着减低,但与健康对照组比较则显着较高,差异具有统计学含义(P<0.05)。见表2。

2.3 免疫组化成果

术中留取患者乳腺癌安排以及癌旁正常安排进行免疫组化染色,咱们发现,C-erbB-2、p-Akt蛋白在患者乳腺癌安排以及癌旁正常安排中均有表达,但乳腺癌安排中表达水平显着高于癌旁正常安排,差异具有统计学含义(P<0.05)。见封三图1 A~E。

2.4 生计预后剖析

运用Kaplan-Meier法对术后随访患者的5年生计率进行剖析并终究发现,癌安排C-erbB-2、p-Akt双阳性患者中位生计期和5年总生计率显着低于单阳性者和双阴性者,见封三图2。

3 评论

跟着女人乳腺癌患者对日子质量和形体美的要求越来越高,保存乳头乳晕的乳腺癌彻底治愈术的运用越来越广泛[6]。现在临床上一般运用保乳手术联合Ⅰ期或Ⅱ期乳房再造手术进行医治[7]。可是,大部分东方女人乳房体积较小,如约束切除规模则影响手术医治效果,这也是现在保乳手术医治的首要妨碍[8]。探究特异性的肿瘤标志物可能对猜测手术危险以及预后提示具有重要含义[9]。

C-erbB-2基因又称为neu或HER-2基因,是一种细胞原癌基因,在多种肿瘤中其癌基因及其蛋白产品(P185)均有过度表达和扩增[10-11]。对C-erbB-2癌基因蛋白产品P185的病理研讨首要多见于乳腺癌,其效果也较为清晰[12]。现在普遍以为,C-erbB-2蛋白产品的阳性表达可作为判别乳腺癌预后的一个独立目标[13]。本研讨发现,在乳腺癌患者中,癌旁正常安排及乳腺癌安排中均可见C-erbB-2蛋白表达,但癌安排中表达水平显着增高,这一成果与既往研讨较为类似,在多种女人有关的肿瘤中,均可见C-erbB-2表达反常,包含子宫内膜癌[14]、卵巢癌[10]以及乳腺癌[15]等。此外,C-erbB-2也能够经过与胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)联合效果一起調控卵巢癌的发作开展[10]。有研讨成果提示,乳腺癌的发作可能与C-erbB-2基因反常表达有关[16]。

p-Akt是磷酸化的Akt,既往报导Akt信号通路在多种癌症疾病中均有表达,如肺癌[17]、胃癌[18]等常见肿瘤。现在,Akt及其信号通路是基础研讨和药物研制范畴中最抢手的激酶和激酶通路之一[19]。PI3K/Akt信号通路关于细胞增殖、分解和凋亡的调理是必要的,其活化与肿瘤发作及肿瘤侵袭搬运密切相关[20]。在Hela细胞中,有研讨发现,干扰素β能够经过NF-κB/p-Akt的信号通路有用促进低剂量顺铂的医治效果[21]。本研讨发现,在乳腺导管癌患者中,p-Akt蛋白在健康乳腺安排、癌安排和癌旁正常安排中均有表达,但在癌安排中表达水平显着高于正常安排,差异具有统计学含义(P<0.05)。这一成果表明,Akt信号通路在乳腺安排癌变的过程中也发挥了重要的效果。

此外,运用Kaplan-Meier剖析本组乳腺癌患者生计状况后发现,C-erbB-2和p-Akt双阳性的乳腺癌患者5年生计率显着低于单阳性者与双阴性的乳腺癌患者,差异具有统计学含义(P<0.05)。咱们剖析,C-erbB-2和p-Akt高表达可能提示乳腺癌患者体内的癌安排处于旺盛的增殖状况。此外,因为保存NAC乳腺癌手术的切除规模较为有限[23],在这部分患者中可能存在潜在的乳腺癌搬运灶,影响了患者的临床预后。因而,咱们以为,关于外周血检测C-erbB-2和p-Akt表达水平增高的非滋润性乳腺癌患者而言,并不合适进行保存乳头乳晕复合体的医治办法。

综上所述,乳腺癌患者外周血C-erbB-2、p-Akt水平显着高于健康对照组,检测C-erbB-2、p-Akt的表达水平对患者承受保存NAC乳腺癌术后的总生计期有重要的临床含义。

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(收稿日期:2017-02-24)

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