【作者】 王海涛；

【导师】 钱海鑫；

【作者基本信息】 苏州大学 ， 普外科， 2010， 博士

【摘要】 结肠癌是我国最常见的恶性肿瘤之一。据2005年第四届全国结直肠癌手术研讨会提供的数据显示:我国结直肠癌发病率已继肺癌、胃癌之后列第三位,占消化道肿瘤的第二位,且近年来发病率呈上升趋势。临床研究分析淋巴结转移是结肠癌最重要的独立预后因素,由于临床Ⅰ、Ⅱ期(AJCC)病例尚缺乏全身辅助化疗对延长生存期有利的证据,故通常未进行化疗。然而却有约20%～30%的局限于肠壁、“无转移”的病例,虽然施行了标准的结肠癌根治术,仍然于5年内不可避免的死于复发或远处转移。根据临床以及基础研究认为最有可能的解释是因为微转移的未能检测,将临床Ⅲ期病例降低分期为Ⅰ、Ⅱ期而未行包括化疗在内的进一步治疗。肿瘤分期是结肠癌最重要的预后因素,与淋巴结转移及数量密切相关,为准确分期,常规检测标本内所有的淋巴结是不切实际的。临床超过69%的转移淋巴结<0.5cm,传统淋巴结检查极有可能遗失包含转移的淋巴结;且淋巴结微转移极易漏诊。这就需要一种新的技术手段,以期尽可能地发现那些隐匿的转移或微转移。前哨淋巴结定位技术(Sentinel lymph node mapping, SLNM)就是一种试图解决以上疑问而提出的且已逐步在临床开展的新技术。前哨淋巴结定位技术是集中对有限的几枚前哨淋巴结(1～4枚)进行详细检查分析,准确评估区域淋巴结状态而改善分期。前哨淋巴结(sentinel lymph node, SLN)的概念由Cabanas等于1977年在描述阴茎癌淋巴引流时提出,直到1992年Morton等成功应用于黑色素瘤以后才在实体瘤中开始了广泛研究。前哨淋巴结被定义为最先直接接受原发肿瘤淋巴引流和首先发生微转移的淋巴结。根据前哨淋巴结观点,肿瘤细胞发生转移是从原发肿瘤为起点,循淋巴管按一定顺序序贯转移的,而最先接受淋巴引流的部位,最有可能发生转移或微转移,并能准确预测区域淋巴结状态。结肠癌淋巴引流系统相对稳定,手术方式较为固定,对结肠癌前哨淋巴结进行定位研究更加能够深入地发现其在消化道实体肿瘤中的应用价值,从而相应地应用于其它消化道肿瘤的诊治。作为一种微创技术,前哨淋巴结定位在黑色素瘤和乳腺癌中的可行性和准确性已得到确认,其成功率高,准确性好,有较好的经济-时间-效益比,可明显提高淋巴结转移诊断率、。但前哨淋巴结在结肠癌中的研究于2000年后才在临床得到发展,其临床作用还存在争议。这就需要进一步研究分析前哨淋巴结定位技术在结肠癌中的真正价值,同时寻求一种在临床上简单易行的操作方法及评价标准。本课题将围绕结肠癌前哨淋巴结定位技术进行基础研究以及临床可行性研究,包括三部分:①结肠癌前哨淋巴结术中定位的可行性及临床价值;②结肠癌前哨淋巴结微转移的检测及其与患者预后的关系;③结肠癌前哨淋巴结微转移与肿瘤病理特性以及肿瘤相关基因的关系。第一部分结肠癌前哨淋巴结术中定位的可行性及临床价值目的:对结肠癌患者进行术中亚甲蓝前哨淋巴结定位(SLNM),分析术中定位前哨淋巴结技术的可行性以及临床价值。方法:对86例结肠癌患者进行术中亚甲蓝(1%)前哨淋巴结定位研究其可行性;术后对前哨淋巴结进行常规苏木精-伊红(HE)染色检测以及免疫组织化学(IHC)检测,分析前哨淋巴结的临床价值。结果:①86例患者术中有82例应用亚甲蓝定位检出SLN,定位成功率为95.3% (82/86)。在82例前哨淋巴结定位成功患者中未发现有异常淋巴引流。②对82例患者全部淋巴结(TLN)进行常规HE检测,共发现35例患者淋巴结转移;通过对SLN进行IHC检测后,共发现48例患者淋巴结转移。有3例患者HE检测示非前哨淋巴结的区域淋巴结(NSLN)有转移,但对SLN检测未发现有SLN转移,SLN检测结肠癌淋巴结转移的假阴性率为8.6%(3/35)。通过对SLN检测,发现常规HE检测无淋巴结转移但SLN检测有淋巴结转移患者13例,使患者由Ⅰ或Ⅱ期提升为Ⅲ期。SLN对结肠癌患者淋巴结转移的诊断敏感率为93.8%(45/48);特异性为91.9%(34/37),SLN诊断准确率为96.3%(79/82)。③82例患者共检出淋巴结1377枚,其中SLN182枚,每例患者SLN 1～4枚不等,平均(2.0±0.49)枚。SLN数占总淋巴结数的13.2% (182/1377)。④182枚SLN中有136枚为肠旁淋巴结,35枚为中间淋巴结,11枚为中央淋巴结。有8例患者发现有跳跃性转移,跳跃性转移发生率为9.8% (8/82),发生跳跃性转移的SLN占SLN总数的11.5%(21/97)。⑤对82例患者淋巴结仅行常规HE检测,发现转移淋巴结共76枚,淋巴结转移率为5.5%(76/1377);转移SLN 43枚,SLN转移率为23.6%(43/182);非SLN的区域淋巴结(NSLN)转移率为2.8%(33/1195),SLN转移率明显高于TLN转移率以及NSLN转移率(P<0.01)。35例有淋巴结转移患者共检出转移淋巴结76枚,其中转移SLN 43枚。常规HE检测有淋巴结转移患者TLN转移率为12.6%(76/602),SLN转移率为54.4%(43/79);NSLN转移率6.3%(33/523)。SLN转移率明显高于TLN转移率和NSLN转移率(P<0.01)。⑥对82例患者淋巴结进行HE检测并对SLN进行IHC检测,共发现转移淋巴结107枚,TLN转移率为7.8%(107/1377);其中转移SLN74枚,SLN转移率为40.7%(74/182);NSLN转移率为2.8%(33/1195)。SLN转移率明显高于TLN转移率和NSLN转移率(P<0.01)。48例有淋巴结转移患者共检出转移淋巴结107枚,其中转移SLN 74枚。48例有淋巴结转移患者TLN转移率为13.0%(107/820),SLN转移率为67.9%(74/109);NSLN转移率4.6%(33/711)。SLN转移率明显高于TLN转移率和NSLN转移率(P<0.01)。结论:①结肠癌患者术中亚甲蓝定位SLN是安全可行的;②结肠癌术中SLN定位能够发现跳跃性转移,但未发现异常淋巴引流;③SLN能够预测结肠癌患者淋巴结转移情况;④SLN能够提高部分结肠癌患者的肿瘤分期。第二部分结肠癌前哨淋巴结微转移的检测及其与患者预后的关系目的:对结肠癌前哨淋巴结进行微转移检测,评价微转移的检测方法及分析前哨淋巴结微转移与患者预后的关系。方法:①对患者前哨淋巴结进行Envision法免疫组织化学(IHC)检测细胞角蛋白20(CK20)表达,②建立实时荧光定量RT-PCR(real-time RT-PCR)检测方法,使用LightCycler荧光定量PCR仪检测前哨淋巴结CK20mRNA表达水平,以GAPDH作为内参。以同一份标本(CK20mRNA拷贝数/GAPDH拷贝数)×104扩来计算CK20mRNA表达指数。③对患者进行随访。④均数之间的比较使用t检验,率的比较采用x2检验或Fisher确切概率法;两组随机计量资料间比较采用非参数检验(Mann-Whitney Test)。两组生存率之间的比较使用Log-rank检验。结果:①对47例常规HE检测无淋巴结转移患者(pN0)进行SLN IHC检测,共发现16枚SLN CK20阳性,有13例患者(13/47,27.7%)SLN CK20阳性。②对47例常规HE检测无淋巴结转移的患者(pN0)进行real-time RT-PCR检测,共发现25例(25/47,53.2%)患者SLN CK20mRNA阳性,共检出CK20mRNA阳性淋巴结31枚;其中有13例患者IHC检测CK20阳性且real-time RT-PCR检测CK20mRNA阳性,有12例(12/47,25.5%)患者IHC检测CK20阴性但real-time RT-PCR检测CK20mRNA阳性。real-time RT-PCR检测微转移敏感性明显高于IHC检测(P<0.01)。13例IHC检测CK20表达阳性患者CK20表达指数(CK20N)范围为37.2～521.9,中位数为247.8;而IHC检测CK20阴性但real-time RT-PCR检测CK20mRNA阳性的12例患者CK20表达指数(CK20N)范围为6.5～134.8中位数为57.8,两组患者CK20mRNA表达指数有明显差异(P<0.001)。③47例常规HE检测淋巴结转移阴性的患者(pN0)有13例(13/47, 27.7%)转移或复发,7例(7/47, 14.9%)死亡;其中IHC检测及real-time RT-PCR检测均为CK20阳性13例患者中5例(5/13, 38.5%)转移或复发,3例(3/13,23.1%)死亡;IHC检测SLN CK20阴性但real-time RT-PCR检测SLN CK20mRNA阳性的12例患者中有4例(4/12,33.3%)转移或复发,2例(2/12, 16.6%)死亡;IHC检测及real-time RT-PCR检测SLN CK20阴性的22例患者中有4例转移或复发(4/22,18.9%),2例(2/22, 9.1%)死亡。有SLN微转移的患者累积复发(转移)风险明显高于无SLN微转移的患者(P<0.05);有SLN微转移的患者累积生存率明显低于无SLN微转移的患者(P<0.05)。结论:①免疫组织化学检测、实时荧光定量RT-PCR检测是发现SLN微转移的灵敏检测方法。②实时荧光定量RT-PCR检测SLN微转移较免疫组织化学检测更敏感。③SLN微转移与患者预后有明显相关性,微转移阳性的患者有较高的复发(转移)机率,相应死亡率也增高。④SLN微转移检测是切实可行的检测方法,可以提高患者淋巴结微转移的阳性率,提高患者的病理分期。第三部分结肠癌前哨淋巴结微转移与肿瘤病理特性以及肿瘤相关基因的关系目的:探讨结肠癌前哨淋巴结微转移与肿瘤病理特性以及肿瘤相关基因的关系。方法:对47例常规HE检测无淋巴结转移的患者(pN0)进行微转移的检测,并对肿瘤进行Envision法免疫组织化学检测其肿瘤相关基因(VEFG、P53、nm23及COX-2)。结果:①对常规HE检测无淋巴结转移的47例患者(pN0)进行SLN微转移的分析后,共发现有25例患者有SLN微转移。②47例患者中,肿瘤最大直径≤2cm患者SLN微转移阳性率为33.3%;肿瘤最大直径>2cm患者SLN微转移阳性率为57.9%,肿瘤最大直径>2cm患者SLN微转移阳性率明显高于肿瘤最大直径≤2cm患者(P<0.05);肿瘤呈肿块型患者SLN微转移阳性率33.3%;溃疡型患者SLN微转移阳性率52.6%;浸润型患者SLN微转移阳性率63.2.%;溃疡型及浸润型患者SLN微转移阳性率明显高于肿块型患者(P<0.05);肿瘤细胞呈高度分化患者SLN微转移阳性率40.0%,中度分化患者SLN微转移阳性率52.6%,低度分化患者SLN阳性率56.5%;数据显示:患者肿瘤细胞分化程度越差,SLN微转移机率越高,但统计学分析未有明显差异((P>0.05);肿瘤浸润至肠壁粘膜或粘膜下层(pT1)患者未及SLN微转移,肿瘤浸润至肠壁固有肌层(pT2)患者SLN微转移阳性率33.3%,肿瘤穿透肌层并至浆膜下(pT3)患者SLN微转移阳性率61.1%,肿瘤穿透浆膜或侵及其他脏器或组织(pT4)患者微转移阳性率为64.7%,随着肿瘤浸润肠壁程度的增加,患者SLN微转移的阳性率逐渐增加(P<0.05)。③47例患者中,VEGF表达阴性患者SLN微转移阳性率为32.0%,VEGF表达阳性患者SLN微转移阳性率为77.3%,VEGF表达阳性患者SLN微转移阳性率明显高于VEGF表达阴性患者(P<0.01);P53表达阴性患者SLN微转移阳性率为47.8%;而P53表达阳性患者SLN微转移阳性率为57.7%,P53表达阳性患者SLN微转移阳性高于表达P53阴性患者,但统计学分析未有明显差异(P>0.05)。nm23表达阴性患者SLN微转移阳性率为60.0%,nm23表达阳性患者SLN微转移阳性率为41.2%,nm23表达阴性患者SLN微转移阳性率明显高于nm23表达阳性患者(P<0.05);COX-2表达阴性患者SLN微转移阳性率为47.6%; COX-2表达阳性SLN微转移阳性率为57.7%,COX-2表达阳性患者SLN微转移阳性率高于COX-2表达阴性患者,但统计学未有明显差异((P>0.05)。结论:①肿瘤病理特性与结肠癌患者的SLN微转移有关;其中肿瘤肿块大体类型、肿瘤最大直径以及肿瘤浸润深度与SLN微转移有明显相关性;②肿瘤相关基因与结肠癌患者的SLN微转移有关,其中VEGF及nm23与SLN微转移有明显相关性。更多还原

【Abstract】 Colon cancer is one of the most common cancers in our country. According to the data of the 2005 fourth nationwide seminar on colorectal operation, the incidence of colorectal cancer is increasing gradually and lies third in all of cancer, only second to lung cancer and stomach cancer. Clinical research indicates that lymph node metastasis is the most important factor affecting its prognosis. Due to absence of favorable evidence to survival, chemotherapy is commonly not applied toⅠandⅡstage colon cancer. However, there were twenty to thirty percent of patients with earlier cancer, which locates within intestinal wall and haD no metastasis, inevitably died of recurrence and distal metastasis though the standard radical operation was enforced on them. According to the clinical and basic research, the most possible factor is the no-check of metastasis, which degrades someⅢstage cancers toⅠorⅡstage cancers, thus chemotherapy was not used. Tumor staging is the most important survival factor, which is closely associated with lymph node metastasis and lymph node quantity. For accurate staging, it is impractical to routinely check all of the lymph nodes. There are more that sixty-nine percent of lymph node smaller than zero point five centimeter of diameter. Traditional surgical check is most likely to neglect lymph node with metastasis. Moreover, micrometastasis is very susceptible to be missed. All of these questions need a new method to find those occult metastases or micrometastases as far as possible. Sentinel lymph node mapping (SLNM) is the new method raised to try to resolve the above questions and now is gradually carried out in clinical. SLNM only needs to focus on some limited lymph nodes (one to four), so it can check them in detail and it can amend the tumor stating accordingly. The conception of SLN was raised in 1977 when Cabanas described the penial lymphatic drainage. It has been widely studied and applied clinically in the solid tumor until Morton successfully applied it to melanoma in 2000. The SLN is defined as the node that receive the tumor’s primary lymphatic drainage at first and that is considered to be most likely to have metastasis or micrometastasis. According to the conception of the SLN, the tumor cell’s metastasis starts from primary tumor, then goes through lymphatic channel sequentially to other site. Thus the site that receives the lymphatic drainage at first is most susceptible to have metastasis and then it can predicate metastasis-situation of regional lymph node. For the lymphatic drainage system of colon is relatively stable and the operation technique is relatively changeless, the SLN mapping to colon cancer is capable to be studied and so we can find its value in the digestive solid tumor and can apply it to the diagnosis and therapy of other solid tumor.The SLNM, as a micro-invasive technique, has a higher diagnosis rate of lymph node metastasis. Its clinical feasibility and accuracy has been validated in the breast cancer and melanoma. It has a good economy-time-effect radio. However the study of the SLNM in colon cancer started from 2000 and its value is still a controversy, which forces us to study its value deeply and try to find an more easily-operated technique and find a good evaluating-standard. Our topic is the basic study and clinical study on the feasibility of the SLNM in colon cancer, which includes three parts:①The clinical feasibility of the SLNM in colon cancer during operation and its clinical value;②The detection of micrometastasis in the SLN of colon cancer and its connection with patients’ prognosis;③the association of SLN micrometastasis with pathologic character and related gene of colon cancer.PartⅠThe clinical feasibility of the sentinel lymph node mapping in colon cancer during operation and its clinical value;Objective: To apply sentinel lymph node mapping(SLNM)to colon cancer during operation with 1% methylence blue and study the its clinical feasibility and its clinical value.Methods: SLNM was implemented with 1% methylence blue during operation on eighty-six patients with colon cancer and the value of SLN was accordingly studied. SLN was checked by means of routine Hematoxylin-Eosin staining(HE) and immunohistochemistry staining(IHC).Results:①SLN was detected in 82 from 86 patients by SLNM with 1% methylence blue during operation and the success rate was 95.3% (82/86).There was no aberrant lymphatic drainage(ALD) to be found among 82 patients.②When only routine HE was applied to check lymph node of 82 patients, there found 35 patients with lymph node metastasis. When IHC was applied further to check SLN, there found 48 patients with lymph node metastasis. There were 3 patients who were diagnosed as lymph node metastasis by HE on none-SLN (NSLN) but diagnosed as no lymph node metastasis by IHC,so the false negative rate of SLN-check was 8.6%(3/35). There found 13 patients who were diagnosed as no lymph node metastasis by HE on SLN but diagnosed as lymph node metastasis by IHC, which upgraded 13 patients from stageⅠorⅡto stageⅢ. The diagnostic sensitivity, specificity and accuracy of SLN is 93.8%(45/48), 91.9%(34/37)and 96.3%(79/82)respectively.③There found 1377 lymph nodes in 82 patients, among them 182 were SLNs. There are 1 to 4 SLNs of one patient and 2.0±0.49 per patient. SLN accounts for 13.2% (182/1377) of total lymph nodes(TLN).④136 among 182 SLNs lied beside the intestinal wall. 35 SLNs were medial lymph nodes and 11 SLNs were central lymph nodes. There were 8 patients had skip metastasis(SM) and skip metastasis rate was 9.8% (8/82).The SLNs of skip metastasis accounted for 11.5% of the total SLNs.⑤When only routine HE was applied to check lymph nodes, there found 76 metastatic lymph nodes and metastatic rate of lymph node was 5.5%(76/1377). There found 43 metastatic SLNs and metastatic rate of SLN was 23.6%(43/182). Metastatic rate of NSLN was 2.8%(33/1195). Metastatic rate of SLN of 82 patients was significantly higher than that of the total lymph node(TLN) and NSLN(P<0.01).In 35 patients who had lymph node metastasis there found 76 metastatic lymph nodes. Among these, 43 were metastatic SLNs. The TLN metastatic rate of the patient who had lymph node metastasis detected only by the routine HE was 12.6%(76/602)while metastatic rate of SLN was 54.4%(43/79 )and metastatic rate of SLN was 6.3%(33/523). Metastatic rate of SLN was significant higher than that of the TLN and NSLN(P<0.01).⑥When the routine HE to lymph node and IHC to SLN of 82 patients, there found 107 metastatic lymph nodes. Among these, there were 74 SLNs. Metastatic rate of TLN was 2.8%(33/1195)metastatic rate of NSLN was 4.6%, while metastatic rate of SLN was 40.7%(74/182).The metastatic rate of SLN was significant higher than that of the TLN and NSLN(P<0.01). There found 107 metastatic lymph nodes in patients who had lymph node metastasis, among these were 74 SLNs. The metastatic rate of TLN was13.0%(107/820)and The metastatic rate of NSLN was 4.6%(33/711),while the metastatic rate of SLN was 67.9%(74/109). The metastatic rate of SLN was significant higher than that of TLN and NSLN(P<0.01).Conclusions:①SLNM during operation on colon cancer with 1% methylence blue was clinically safe and feasibly.②SLNM during operation can find skip metastasis, but we did not find ALD.③SLN can predict the metastatic status of lymph node of colon cancer.④SLN can upgrade some patients’ tumor staging.PartⅡThe detection of micrometastasis in the SLN of colon cancer and its association with patients’ prognosisObjective: To detect the micrometastasis in the SLN of colon cancer and analyze its association with patients’ prognosisMethods:①SLN was detected by means of Envision immunohistochemistry to see if cytokeratin-20 protein was expressed.②Real-time quantitative reverse transcription polymerase chain reaction (real-time RT-PCR) method was established. LightCyler instrument was used for real-time RT-PCR. CK20mRNA expression level of patient’s SLN was detected with GAPDH as inner comparison after operation.CK20mRNA expression index was calculated as CK20mRNA copy/ GAPDH copy×104.③All the patients were followed-up.④T test was used to compare the median. x2 test or Fisher test was used to compare the ratio. Comparison of the random datum of two groups was analyzed with Mann-Whitney test. Comparison of survival rate between two groups was analyzed with Log-rank test.Results:①When SLN was checked by means of IHC in those 47 patients who were diagnosed as no lymph node metastasis (pN0),there were 16 SLNs in which CK20 was positively expressed and 13 patients with CK20 positively expressed in SLN.②When SLN was checked by means of real-time RT-PCR, there found 25 patients of no lymph node metastasis and there were 31 SLNs in which CK20 mRNA was positively expressed and 25 patients with CK20 mRNA positively expressed in SLN. Among those 25 patients, there were 13 patients in whom CK20 is also checked out by IHC and there were 12 patients in which CK20 was only positively detected by real-time RT-PCR. The sensitivity of real-time RT-PCR was significant higher than that of IHC(P<0.01).CK20 expression index ranged from 37.2 to 521. 9 from those 13 patients with positive CK2O expression detected not only by IHC but also by real-time RT-PCR and the median of the CK20 expression index was 247.8. However CK20 expression index ranged from 6.5 to 134.8 in those 12 patients with positive CK2O expression only detected by real-time RT-PCR and the median was 57.8. CK20 expression index of patients with CK20 detected by IHC is significant higher than that of patients detected only by real-time RT-PCR(P<0.001).③Among 47 patients who were diagnosed as no lymph node metastasis by routine HE, there were 13 (13/47, 27.7%)patients who recurred or had metastasis and 7 (7/47, 14.9%)patients died. Among 13 patients who were diagnosed as SLN micrometastasis by IHC ,there were 5(5/13, 38.5%)patients who recurred or have metastasis and 3( 3/13,23.1% ) patients died. Among 12 patients who were diagnosed as SLN micrometastasis not by IHC but by real-time RT-PCR ,there were 5(4/12,33.3%)patients who recurred or had metastasis and 2(2/12, 16.6%)patients died. Among 22 patients who were diagnosed as no SLN micrometastasis by IHC and real-time RT-PCR ,there were 4(4/22,18.9%)patients who recurred or had metastasis and 2(2/22, 9.1%)patients died.The cumulative recurrence(metastasis) rate of patients with SLN micrometastasis was higher than that of patients without SLN micrometastasis(P<0.05).The cumulative survival rate of patients with SLN micrometastasis was lower than that of patients without SLN micrometastasis.Conclusions:①IHC and real-time RT-PCR are sensitive means to detect SLN micrometastasis.②Real-time RT-PCR is more sensitive than IHC to detect SLN micrometastasis.③SLN micrometastasis is closely associated with patients’ prognosis. There is a higher recurrence rate and metastasis rate in the patients with SLN micrometastasis.④The detection of SLN micrometastasis is a clinically feasible technique, which can raise the positive rate of lymph node then upgrade the patient’s pathological staging.PartⅢthe association of SLN micrometastasis with the pathologic character and related gene of colon cancer.Objective: to study the association of SLN micrometastasis with the tumor pathologic character and tumor related gene in colon cancer.Methods: SLN micrometastasis was detected in 47 patients who were checked as no lymph node metastasis(pN0). The related genes (VEFG、P53、nm23及COX-2) were checked by means of IHC.Results:①There were 25 patients with SLN micrometastasis among 47 patients who were detected as no lymph node metastasis(pN0).②SLN micrometastasis rate of the patient whose tumor diameter was no less than 2 centimeters was 57.9%, while that of the patient whose tumor diameter was less than 2 centimeters was 33.3%.The former was significant higher than the latter(P<0.05). SLN micrometastasis rates of the patients who had mass-type, ulcerated-type and infiltrating-type tumor were 33.3%, 52.6% and 63.2% respectively. The former two were significant higher than the latter one (P<0.05). SLN micrometastasis rates of the patients whose tumor cells were well, medially and poorly differentiated were 40.0%, 52.6% and 56.5% respectively. As the tumor cell gradually poorly differentiated, SLN micrometastasis rates of the patients gradually increased. But there was no statistical difference between their SLN micrometastasis rate(P>0.05).There was no SLN micrometastasis in patients whose tumor was classified as pT1. SLN micrometastasis rates of patients whose tumors were classified as pT2, pT3 and pT4 were 33.3%, 61.1% and 64.7% respectively. As the tumor infiltrated deeper into intestinal wall, the patient’ SLN micrometastasis rate increased gradually(P < 0.05).③SLN micrometastasis rate of patient whose tumor negatively expressed VEGF was 32.0%, while that of patient whose tumor positively expressed VEGF was 77.3%. The latter was significantly higher than the former(P<0.01). SLN micrometastasis rate of patient whose tumor negatively expressed P53 was 47.8%, while that of patient whose tumor positively expressed P53 was 57.7%. The latter was higher than the former, but there is no statistical difference(P > 0.05). SLN micrometastasis rate of patient whose tumor negatively expressed nm23 was 60.0%, while that of patient whose tumor positively expressed nm23 was 41.2%. The former was significantly higher than the latter(P < 0.05). SLN micrometastasis rate of patient whose tumor negatively expressed COX-2 was 47.6%, while that of patient whose tumor positively expressed COX-2 was 57.7%. The latter was higher than the former, but there is no statistical difference(P>0.05).Conclusions:①Tumor pathologic character is associated with SLN micrometastasis. Among tumor pathologic characters, configuration, diameter and infiltrating depth are closely associated with SLN micrometastasis.②Some tumor genes are associated with SLN micrometastasis. Among tumor genes, VEGF and nm23 are closely associated with SLN micrometastasis.更多还原