| Objectives This study was designed to detect the expression level of BCL-6?CDC7 in peripheral blood plasma free DNA and pathological tissues of Diffuse large B-cell lymphoma(DLBCL)patients. And to evaluate the expression level in DLBCL early diagnosis, treatment evaluation and prognosis of clinical significance.Methods Collected and analyzed the clinical and pathological data of 54 cases DLBCL patients in Shandong Cancer Hospital and affiliated hospital of Shandong Province Academy of Medical Sciences from March 2013 to June 2015. Plasma of free DNA samples were extracted in 20 primary ?20 relapse and14 Clinical remission cases of DLBCL, and 20 healthy controls. To detect the Plasma of free DNA expression levels in each group. The expression of BCL-6 and CDC7 in plasma free DNA was detected by PCR method. Collection of paraffin embedded tissue specimens from all patients before tumor treatment, the expression of BCL-6 and CDC7 in paraffin embedded tissues was detected by Immunohistochemical method. Statistics survival data, compared the survival rate of patients with different BCL-6 and CDC7 expression. Comparison between the two groups rate using the?2 test rate; OS and PFS were analyzed with Kaplan-Meier survival curves,between the two groups by Log-rank test; multivariate analysis using Cox regression model. Using SPSS17.0 software package for statistical analysis, test level: Bilateral? = 0.05, P <0.05 was considered statistically significant.Results 1.Plasma free DNA was successfully obtained from 54 cases of DLBCL patients and healthy controls. Determination of plasma free DNA concentration by UV spectrophotometry, the DNA concentration in incipient group?refractory group?remission group ?control group respectively were 120.6?24.0 ng/ml?117.0?27.6 ng/ml?2.5?0.8 ng/ml?2.2?0.8ng/ml?Incipient group and refractory group DNA was significantly higher than remission group, the difference was statistically significant.2.In plasma free DNA detection, BCL-6 positive expression was 55.6%, negative expression was 44.4%;2-years overall survival of the patient with BCL-6 positive expression and negative expression were76.6% ?62.5%(P =0.009);2-years progress free survival of the patient with BCL-6 positive expression and negative expression were46.6%?33.3%(P =0.246);BCL-6 expression correlated to clinical stage and IPI scores(P<0.05)?CDC7positive expression was51.9%, negative expression was 48.1%;2-years overall survival of the patient with CDC7 positive expression and negative expression were40.4%?66.7%(P =0.045);2-years progress free survival of the patient with CDC7 positive expression and negative expression were28.7%?46.8%(P=0.048);CDC7expression correlated to clinical stage ?IPI scores and Term effect(P<0.05)?3.In immunohistochemistry experiment, BCL-6 positive expression was 61.1%, negative expression was38.9%;2-years overall survival of the patient with BCL-6 positive expression and negative expression were72.7% ?61.9%(P =0.027);2-years progress free survival of the patient with BCL-6 positive expression and negative expression were51.5%?38.1(P =0.372);BCL-6 expression correlated to clinical stage and IPI scores(P<0.05)?CDC7positive expression was46.3%, negative expression was 53.7%;2-years overall survival of the patient with CDC7 positive expression and negative expression were44.0%?75.9%(P =0.034);2-years progress free survival of the patient with CDC7 positive expression and negative expression were32.0%?48.3%(P =0.108);CDC7expression correlated to clinical stage ?IPI scores ?LDH and term effect(P<0.05)?4.BCL-6 expression in plasma DNA and pathological tissues were 55.6% and 61.1%, P = 0.558; CDC7 expression in plasma DNA and pathological tissues were 51.9% and 46.3%, P =0.564, all P > 0.05, the difference was not statistically significant.Conclusion 1.Plasma free DNA concentration DLBCL patients was significantly higher than the level of healthy people, there is a significant difference compared with healthy people; Free DNA levels in complete remission DLBCL patients was nearly healthy people level, the difference was not statistically significant. Free DNA concentration detection has a certain significance in the early diagnosis and guide treatment of DLBCL patients.2.BCL-6 expression correlated to clinical stage and PIP score and 2-year OS, whereas no significant relationship with the patient's gender, age, B symptoms, term effect and 2-year PFS. CDC7 expression correlated to clinical stage and IPI score and 2-year OS, whereas no significant relationship with the patient's gender, age, B symptoms, the efficacy and 2-year PFS.CDC7 expression and clinical stage?IPI score,term effect and 2-year OS related, whereas no significant relationship with the patient's gender, age and B symptoms and 2-years progress free survival.3.BCL-6, CDC7 in plasma free DNA detection and immunohistochemical detection in judging the prognosis of DLBCL patients has consistency. DLBCL patients BCL-6 and CDC7 in plasma DNA and pathological tissue levels were not significantly different, and therefore the detection of plasma free DNA is expected to become a new index DLBCL detection. |
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