| Objective: Cytokeratin 19, as an indicator of the detection of circulating tumor cells indicators, gradually attracts attention. The present study was designed to use reverse transcription polymerase chain reaction(RT-PCR) to detect the expression levels of CK19 m RNA in peripheral blood of gastrointestinal cancer patients in stage II and III, and to investigate the prognostic value of CK19 m RNA in the peripheral blood of gastrointestinal cancer patients in stage II and III for clinical guide.Methods: A sample of 133 gastrointestinal cancers was included which had radical surgery for TNM stage II and III by pathological and hospitalized in 2013.05-2014.01. Every 10 ml peripheral blood sample was obtained from the peripheral blood of the patients.All blood samples were obtained at the middle of vein puncture after the first 5 m L of blood were discarded, and the other 5 ml was kept in EDTA tube and processed. Blood cell lysing reagents(Invitrogen) removed red blood cells. Cell pellets were collected and Total RNA was extracted. The amplification of RNA was checked by RT-PCR. Then standard curve was made. The expression of CK19 m RNA was detected using this method in the peripheral blood of the patients. The treatment of patients was not interfered, and an assessment of tumor for patients was taken every three months until disease relapse or death of patients was followed up with. We analysis the relationship of CK19 m RNA and clinical pathological features such as TNM staging, sex, age, and lymph node metastasis. Statistical analysis: all data were completed using SPSS19.0 software, data expressed as a percentage, or variance. Kaplan-Meyer survival curves method, univariate survival is estimated by log-rank, Cox proportional hazards model was used for multivariate survival analysis. P <0.05 was statistically significant.Results: 1. This study includes statistics of 133 gastrointestinal cancer, which had radical surgery.The number of male and female are 99 and 34, the rate is 2.91:1.The onset age is 27-83, the median age is 61.08±9.88. 2. Of all the 133 gastrointestinal cancer, 71( 53.4%) are in the gastric cancer and CK19 m RNA+ were detected in 25(35.2%); while 42(31.6%) in colorectal cancer, CK19 m RNA+ in 11(26.2%); 20(15%) in esophageal cancer, CK19 m RNA+ in 5(3.7%). 3. Of all the 133 gastrointestinal cancer, 71( 53.4%) are in stage II for TNM and CK19 m RNA+ were detected in 24(33.8%); while 62( 46.6%) are in stage III for TNM and CK19 m RNA+ were detected in 17(27.4%). 4. We followed up 133 cases, the follow-up rate was 100%.Of them, 46(34.6%) had distant metastasis and CK19 m RNA+ was detected in 32(69.6%); 8(6.0%) died and CK19 m RNA+ was detected in 2(25%). 5. CK19 m RNA+ was detected in 30.8% of the 133 patients. Kaplan-Meier analysis also showed the detection of CK19 m RNA+ cells was associated with shorter disease free survival(P<0.05). Cox proportional hazard models indicated that the detection of CK19 m RNA+ was an independent prognostic factor. The expression of CK19 m RNA in the peripheral blood was no relationship to gander, age, histology grade, lymph nodes metastasis(P>0.05).Conclusion: A part of patients with gastrointestinal cancer in the early stage have micrometastases in the peripheral blood. The detection of CK19 m RNA+ cells expression in the peripheral blood predicts poor prognosis(short DFS) in patients with early gastrointestinal cancer. CK19 m RNA, as a genetic markers in detection of micrometastases of early gastrointestinal cancer patients, have a high value, which helps to determine whether early gastrointestinal tumor patients have disease progression and distant metastasis, so as to is estimated to clinicians early intervention, extend the prognosis of patients. |
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