血清MicroRNA作为结直肠癌肝转移患者早期诊断标志物的探索性研究

Serum MicroRNA as a marker for early diagnosis in patients with colorectal liver metastases

目的通过对结直肠癌患者血清进行实时定量PCR检测,筛选出作为结直肠癌肝转移早期诊断标志物的相关miRNA。方法通过文献检索,筛选出11个转移相关的miRNA(miRs-31,335,206,141,126,200b,200c.21,Let7a,Let 7b和Let 7c),收集2007年7月至2013年4月在北京友谊医院收治的术前留置血清的结直肠癌转移(IV期)病例共计108例,其中结直肠肝转移患者72例(SLM-CRC),其他脏器转移患者36例(OM-CRC);并纳入2008年1月至2010年6月术前留置血清的结直肠癌(L-CRC)(Ⅰ-Ⅲ期)病例共计116例作为对照组进行研究。通过对各亚组血清样本进行实时定量PCR检测结果找出结直肠癌肝转移相关miRNA。结果筛选出11个转移相关的miRNA中的7个miRNA(miRs-31、141、126、21、Let7a、Let7b和Let7c)可以从血清中被检测到。与局灶性结直肠癌患者相比,miR-141、miR-126、Let-7a和miR-21在转移性结直肠癌患者的血清中表达具有显著性差异(P<0.0001,P<0.0001,P=0.0120和P<0.0001)可以作为结直肠癌转移相关MiRNAs;miR-141,miR-126和miR-21在结直肠癌同时性肝转移患者血清中表达具有显著性差异(P<0.0001,P<0.0001和P<0.0001)可以作为结直肠癌肝转移相关MiRNAs。ROC曲线下miR-126有一个截点值(log_(10)^(RQ)=-0.2005)的77.78%的敏感度、68.97%的特异性;miR-141有一个截点值(log_(10)^(RQ)=-0.2285)的86.11%的敏感度、76.11%的特异性;miR-21有一个截点值(log_(10)^(RQ)=-0.1310)的73.61%的敏感度、66.38%的特异性;miR-141与miR-126、miR-21相比有更好的敏感度及特异性。结论 miR-141、miR-126和miR-21是结直肠癌肝转移相关MiRNAs,其中miR-141有更好的灵敏度及特异性,进一步证实miR-141可以作为结直肠癌肝转移患者早期诊断标志物。

Objective To investigate the distribution of the difference in the expression of miRNAs in patients with colorectal liver metastases.Methods We reviewed the literature in search for studies reporting on miRNA expression profiles of motile or invasive cells and obtained 11 miRNAs which were related with metastasis in colorectal cancer（CRC）（miRs-31,335,206,141,126,200 b,200c,21,Let7 a,Let7b and Let 7c）.Serum samples from CRC patients that had been surgically treated were obtained from Beijing Friendship Hospital,including 116 consecutive patients of localized CRC（L- CRC）（stage I-Ⅲ） between January 2008 and June 2010,and 108 consecutive patients of metastatic CRC,including 72 with synchronous liver metastasis（SLM- CRC） and 36 with other organ- metastasis（OM- CRC）,between September 2007 and April 2013.The miRNAs expressions in serum were analyzed by real- time quantitative PCR in all subgroups of CRC patients.Results Seven（miRs-31,141,126,21,Let7 a,Let7b and Let7c） of 11 tested miRNAs could be detected from serum.Four miRNAs,miR- 126,miR-141,Let- 7a and miR- 21 were identified as metastasis- associated miRNAs（P〈0.0001,P〈0.0001,P = 0.0120 and P〈0.0001,respectively）.Three miRNAs,miR- 126,miR- 141 and miR- 21 were identified as liver metastasis- associated miRNAs（P〈0.0001,P〈0.0001 and P〈0.0001,respectively）.The receiver operating characteristic curve showed serum miR- 126 had a cut- off（log10^RQ =-0.2005） with77.78%sensitivity and 68.97%specificity with an area under curve of 0.7564,miR-141 had a cut- off（log10^RQ=-0.2285） with 86.11%sensitivity and 76.11%specificity with an area under curve of 0.8279,miR-21 had a cut- off（log10^RQ =-0.1310） with 73.61%sensitivity and 66.38%specificity with an area under curve of 0.7479.Compared with miR-126 and miR- 21,miR- 141 had a better sensitivity and specificity.Conclusion miR- 141,miR- 126 and miR-21 were related to colorectal liver metastasis.miR- 141 has excellent sensitivity and specificity to be used as an early diagnostic marker in colorectal cancer with liver metastases.