摘要目的 探讨同时性结直肠癌肝转移(SCRLM)患者结直肠癌组织中Kiss-1蛋白和MMP9蛋白的表达及其与SCRLM患者临床病理因素和预后的关系,以及两者表达的相关性.方法 采用回顾性病例对照研究方法.收集2000年1月至2013年5月第二军医大学附属东方肝胆外科医院收治的96例SCRLM患者和69例无转移结直肠癌患者的临床病理资料.收集96例SCRLM患者手术切除的结直肠癌组织及其中50例癌旁组织(距离阴性切缘≥5 cm);69例无转移结直肠癌患者手术切除的结直肠癌组织进行研究.采用免疫组织化学染色检测Kiss-Ⅰ蛋白和MMP9蛋白的表达情况.通过门诊和电话方式随访患者的生存情况,随访时间截至2014年8月.计数资料的比较及SCRLM患者结直肠癌组织中Kiss-1蛋白和MMP9蛋白表达与患者临床病理因素的关系采用x2检验或Fisher确切概率法.采用Kaplan-Meier法绘制生存曲线,采用Log-rank检验进行生存分析.相关性分析采用Pearson等级相关分析.结果 Kiss-1蛋白的表达位于组织细胞的细胞质,其阳性表达率在SCRLM患者结直肠癌组织、无转移结直肠癌患者的结直肠癌组织、SCRLM患者结直肠癌旁组织中分别为24.0％(23/96)、43.5％(30/69)、52.0％ (26/50),3者比较,差异有统计学意义(x2=14.307,P＜0.05).其中SCRLM患者结直肠癌组织中Kiss-1蛋白的阳性表达率分别与无转移结直肠癌患者的结直肠癌组织和SCRLM患者结直肠癌旁组织比较,差异均有统计学意义(x2=0.702,11.594,P＜0.05);无转移结直肠癌患者的结直肠癌组织与SCRLM患者结直肠癌旁组织比较,差异无统计学意义(x2=0.845,P＞0.05).MMP9蛋白的表达位于组织细胞的细胞质,其阳性表达率在SCRLM患者结直肠癌组织、无转移结直肠癌患者的结直肠癌组织、SCRLM患者结直肠癌旁组织中分别为67.7％ (65/96)、62.3％ (43/69)、36.0％ (18/50),3者比较,差异有统计学意义(x2=14.203,P＜0.05).其中SCRLM患者结直肠癌组织中MMP9蛋白的阳性表达率分别与无转移结直肠癌患者的结直肠癌组织和SCRLM患者结直肠癌旁组织比较,差异均有统计学意义(x2=13.475,13.475,P＜0.05);无转移结直肠癌患者的结直肠癌组织中MMP9蛋白的阳性表达率与SCRLM患者结直肠癌旁组织比较,差异无统计学意义(x2=8.038,P＞0.05).SCRLM患者结直肠癌组织中Kiss-1蛋白阳性表达率在肿瘤高、中、低分化中分别为66.7％、21.9％、17.6％,在肿瘤浸润肌层、浆膜外、浆膜层SCRLM患者结直肠癌组织中分别为50.0％、28.6％、17.5％,在无和有淋巴结转移SCRLM患者结直肠癌组织中分别为44.0％、16.9％.不同肿瘤分化程度、肿瘤浸润深度、淋巴结转移情况的SCRLM患者结直肠癌组织中Kiss-1蛋白的表达比较,差异均有统计学意义(x2=6.546,6.172,7.453,P＜o.05).MMP9蛋白阳性表达率在肿瘤浸润肌层、浆膜外、浆膜层SCRLM患者结直肠癌组织中分别为25.0％、66.7％、76.2％,在无和有淋巴结转移SCRLM患者结直肠癌组织中分别为44.0％、76.1％.不同肿瘤浸润深度、淋巴结转移情况的SCRLM患者结直肠癌组织中MMP9蛋白的表达比较,差异均有统计学意义(x2=12.094,8.690,P＜0.05).96例患者均获得术后随访.随访时间为12 ～ 176个月,中位随访时间为68个月.Kiss-1蛋白表达阳性的SCRLM患者中位总体生存时间为31个月,中位无瘤生存时间为26个月,5年累积生存率为69.6％,5年无瘤生存率为26.1％;Kiss-1蛋白表达阴性的SCRLM患者中位总体生存时间为26个月,中位无瘤生存时间为19个月,5年累积生存率为24.7％,5年无瘤生存率为12.3％.两者总体生存情况和无瘤生存情况比较,差异均有统计学意义(x2=16.578,14.436,P＜0.05).MMP9蛋白表达阳性的SCRLM患者中位总体生存时间为31个月,中位无瘤生存时间为19个月,5年累积生存率为24.6％,5年无瘤生存率为12.3％;MMP9蛋白表达阴性的SCRLM患者中位总体生存时间为28个月,中位无瘤生存时间为16个月,5年累积生存率为58.1％,5年无瘤生存率为22.6％.两者总体生存情况和无瘤生存情况比较,差异均有统计学意义(x2=14.073,8.532,P＜0.05).SCRLM患者结直肠癌组织中Kiss-1蛋白表达阳性23例(其中MMP9蛋白表达阳性10例、阴性13例),阴性73例(其中MMP9蛋白表达阳性55例、阴性18例),Kiss-1蛋白和MMP9蛋白表达呈负相关(r=-0.291,P＜0.05).结论 Kiss-1蛋白在SCRLM患者结直肠癌组织中表达降低,MMP9蛋白在SCRLM患者结直肠癌组织中表达升高,这与结直肠癌侵袭、转移、预后密切相关.两者表达呈负相关,提示两者联合检测有望成为预测SCRLM预后的标志物.

abstractsObjective To investigate the expressions and correlations of Kiss-1 and matrix metalloproteinases-9 (MMP9) proteins in the colorectal cancer (CRC) tissues of patients with synchronous colorectal liver metastasis (SCRLM),and the association with the clinicopathologic factors and prognosis of patients.Methods The retrospective case-control study was adopted.The clinicopathological data of 96 patients with SCRLM and 69 patients with CRC and no metastasis who were admitted to the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University from January 2000 to May 2013 were collected.The 96 CRC tissues and 50 adjacent normal tissues (distance from resection margin ≥ 5 cm) were collected from 96 patients with SCRLM,and 69 CRC tissues were collected from 69 patients with CRC and no metastasis.Expressions of Kiss-1 and MMP9 protein were detected by immunohistochemistry (IHC).The follow-up of outpatient examination and telephone interview was performed to detect survival of patients till August 2014.Comparison of count data and correlation between expressions of Kiss-1 or MMP9 protein and clinicopathological factors were analyzed by the chi-square test and Fisher exact probability.Survival curve was drawn using the Kaplan-Meier method,and survival analysis was done using the Log-rank test.Correlation analysis was done by the Pearson correlation.Results Expression of Kiss-1 protein was located in the cytoplasm of tissue cells.The positive expression rates of Kiss-1 protein in CRC tissues of patients with SCRLM and with CRC and no metastasis and in adjacent normal tissues were 24.0％ (23/96),43.5％ (30/69) and 52.0％ (26/50),respectively,with a significant difference among the 3 tissues (x2 =14.307,P ＜ 0.05) and no significant difference between CRC tissues of patients with CRC and no metastasis and patients with SCRLM (x2 =0.845,P ＞ 0.05).The positive expression rate of Kiss-1 protein in CRC tissues of patients with SCRLM was significantly different from that in CRC tissues of patients with CRC and no metastasis and in adjacent normal tissues (x2 =0.702,11.594,P ＜ 0.05).Expression of MMP9 protein was located in the cytoplasm of tissue cells.The positive expression rates of MMP9 protein in CRC tissues of patients with SCRLM and with CRC and no metastasis and in adjacent normal tissues were 67.7 ％ (65/96),62.3 ％ (43/69) and 36.0％ (18/50),respectively,with a significant difference among the 3 tissues (x2=14.203,P ＜0.05) and no significant difference between CRC tissues of patients with CRC and no metastasis and patients with SCRLM (x2=8.038,P ＞ 0.05).The positive expression rate of MMP9 protein in CRC tissues of patients with SCRLM was significantly different from that in CRC tissues of patients with CRC and no metastasis and in adjacent normal tissues (x2 =13.475,13.475,P ＜ 0.05).The positive expression rates of Kiss-1 protein in CRC tissues of patients with SCRLM were 66.7％,21.9％ and 17.6％ in the high-,mederate-and low-differentiated tumor,50.0％,28.6％ and 17.5％ in the muscular layer of tumor invasion,outside of serosa and serosal layer,44.0％ and 16.9％ in patients with and without lymph node metastasis,respectively,showing significant differences among the tumor differentiation degree,depth of tumor invasion and lymph node metastasis (x2=6.546,6.172,7.453,P ＜0.05).The positive expression rates of MMP9 protein in CRC tissues of patients with SCRLM were 25.0％,66.7％ and 76.2％ in the muscular layer of tumor invasion,outside of serosa and serosal layer,44.0％ and 76.1％ in patients with and without lymph node metastasis,respectively,showing significant differences between the depth of tumor invasion and lymph node metastasis (x2 =12.094,8.690,P ＜ 0.05).All the 96 patients with SCRLM were followed up for a median time of 68 months (range,12-176 months).The median overall survival time,median tumor-free survival time,5-year cumulative survival rate and 5-year tumor-free survival rate were 31 months,26 months,69.6％ and 26.1％ in patients with SCRLM and positive expression of Kiss-1 protein and 26 months,19 months,24.7％ and 12.3％ in patients with SCRLM and negative expression of Kiss-1 protein,respectively,showing significant differences between the overall survival and tumor-free survival (x2=16.578,14.436,P ＜ 0.05).The median overall survival time,median tumor-free survival time,5-year cumulative survival rate and 5-year tumor-free survival rate were 31 months,19 months,24.6％ and 12.3％ in patients with SCRLM and positive expression of MMP9 protein and 28 months,16 months,58.1％ and 22.6％ in patients with SCRLM and negative expression of MMP9 protein,respectively,showing significant differences between the overall survival and tumor-free survival (x2=14.073,8.532,P ＜0.05).Of 96 patients with SCRLM,there were 23 patients with positive expression of Kiss-1 protein (10 with positive expression of MMP9 protein and 13 with negative expression of MMP9 protein) and 73 with negative expression of Kiss-1 protein (55 with positive expression of MMP9 protein and 18 with negative expression of MMP9 protein),with a negative correlation between expressions of Kiss-1 protein and MMP9 protein (r =-0.291,P ＜ 0.05).Conclusions The reduced expression of Kiss-1 protein and elevated expression of MMP9 protein are closely associated with invasion and metastasis of CRC and prognosis of patients.A combination detection of Kiss-1 and MMP9 proteins is expected to become a marker for predicting the prognosis of patients with SCRLM based on the negative correlation between them.