Clinical Research On The Coagulation Index Change To Evaluate The Chemotherapeutic Efficacy Against Breast Cancer And The Tumor Metastasis Risk Through Blood Circulation

Various blood coagulation factors secreted by malignant cells may involve in theinvasion and metastasis of tumor. The abnormal blood coagulation index in patients withbreast cancer is probably related with clinical and pathological characteristics of tumor, but itstill remains a controversial issue. Neoadjuvant chemotherapy plays an important role inbreast cancer treatment. What kind of change does the blood coagulation index demonstrateafter chemotherapy? The coagulation index before and after chemotherapy are different. Arethis related with the chemotherapeutic efficacy? Can coagulation index at baseline and afterchemotherapy be regarded as the predictor of chemotherapy? These questions, rarelymentioned in the reference, are currently disputed. Previous studies have suggested that theHCS （hypercoagulable state） caused by malignant tumor may participate in the invasion andmetastasis of tumor cells, but the direct evidence is absent; CTC （Circulating Tumor Cell） isregarded as a sign of breast cancer micrometastasis, which is an important step in distantmetastasis. Is there any correlation between the blood coagulation index change and CTCpositive? This study aimed to explore the above-mentioned issue.Methods&Results1. Preliminary study on the relationship between blood coagulation index andclinical and pathological characteristicsMethod: In158patients with stage Ⅰ～Ⅲ breast cancer,28with stage â…£and94withbreast benign disease,5ml venous blood was taken before treatment to detect the followingindex: PT （Prothrombin Time）, PT%, PTINR（Prothrombin time international normalizedratio，PTINR）,APTT （Activated Partial Thromboplastin Time）, APTT Ratio, Fib （Fibrinogen）, D-D（D-dimer）, TT （Thrombin Time）, PLT（Platelet）. Meanwhile the expressions of ER, PRand HER-2were detected by immunohistochemical method. Then the blood coagulationrelated index were compared between breast cancer with different stage and breast benigndisease, and the correlation between blood coagulation index and tumor size, pathologicalstatus of lymph node, TNM staging, ER, PR, HER-2, molecular subtypes of breast cancer wasanalyzed.Results:1) Compared with breast benign disease, PTINR in patients with breast cancerwas lower[0.94（0.08）vs0.96（0.07），P=0.01]; Fib was higher [（3.00±0.71） vs （2.52±0.44）,P=1.83×e^-11]; D-D was higher [133（140.75） vs104.5（73.5）, P=7.78×e^-5].2) Compared withstage I-III breast cancer, Fib level in stage â…£ was higher (3.49±0.93g/L, P=2.26×e^-4), butthere was no statistical difference among stage I, â…¡,III. Compared with that of group T₁-T₃,the Fib level of group T4was higher（3.66±1.16g/L, P=0.04）,but there was no statisticaldifference among group T₁, T₂, T₃. Compared with that of group N₀-N₂, the Fib level ofgroup N3was higher（3.48±0.92g/L, P=0.04）, but there was no statistical difference amonggroup N₀, N1, N₂. D-D level in ER-negative group was higher than that in ER-positive group[145（176.5） vs116.5（113）, P=0.04]. APTT and TT in the patients with HER-2receptorpositive were longer [（25.4±3.57） vs （23.91±3.51）, P=0.04;（16.83±1.17） vs （16.31±1.28,P=0.04）]; D-D level was increased [158（163） vs116（122）, P=0.03]. The Fib level inBasal-like type breast cancer was higher than that in HER-2type [（3.08±0.56） vs （2.65±0.57）,P=0.04], but no statistical difference was shown between Basal-like and Luminal, HER-2andLuminal. There was no difference of blood coagulation index between ER-positive group andER-negative group.2. Preliminary study on the relationship between the changes of blood coagulationindex before and after Neoadjuvant chemotherapy and the chemotherapeutic efficacyMethod:5ml venous blood was drawn from146patients with breast cancer before andafter neoadjuvant chemotherapy （Paclitaxel175mg/m2or Docetaxel75mg/m2, Epirubicin75mg/m2, d1,21d per cycle）, to detect blood coagulation related index. The change of bloodcoagulation index before and after neoadjuvant chemotherapy, the relationship between bloodcoagulation index and the chemotherapeutic efficacy, the relationship between the change ofblood coagulation index before and after neoadjuvant chemotherapy and the chemotherapeutic efficacy were analyzed.Results:1) Fib level decreased in28patients （19.2%）, remained the same in1patient（0.7%）, increased in117patients （80.1%） after chemotherapy,Fib was significantly increasedfrom （2.82±0.62）g/L before chemotherapy to （3.44±0.78）g/L after chemotherapy （P＜4.162×e-16）. D-D level decreased in92patients （63%）, increased in54patients （37%） afterchemotherapy,D-D was significantly decreased from （162.41±132.03） ng/ml beforechemotherapy to （142.10±13.17） ng/ml after chemotherapy （P＜4.162×e-16）; so the D-D levelshowed a significant difference before and after chemotherapy (P＜2.123×e^-4). PLTdecreased in27patients （18.5%）, increased in119patients （81.5%） after chemotherapy, PLTwas significantly increased from （193.10±55.15）×10⁹/L before chemotherapy to（245.15±77.70）×10⁹/L after chemotherapy, so the PLT had a significant difference （P＜1.586×e-14） before and after chemotherapy. PT, PT%, PTINR, APTT, APTT Ratio, TT leveldid not show notable change before and after chemotherapy.2) The D-D level ofchemotherapy effective group was higher than that of non-effective group （140（125.5） vs94.5（96.75）, P=0.001）. The PT, PT%, PTINR, APTT, APTT Ratio, Fib, TT, PLT level of bothgroups did not show notable change.3) There is no notable change observed of bloodcoagulation index after chemotherapy between effective chemotherapy group andnon-effective chemotherapy group.4) The D-D level was decreased in92patients afterchemotherapy, among them, chemotherapy was effective to61patients （66.30%）,non-effective to31（33.70%）. The D-D level was increased in54patients after chemotherapy,among them, chemotherapy was effective to19patients （35.19%）, non-effective to35（33.70%）. For the those who had an increased D-D, chemotherapy was effective to19（35.19%）, non-effective to35（64.81%）, and the difference was statistically significantbetween the effective and non-effective ones (P＜2.649×e^-4). PT, PT%, PTINR, APTT, APTTRatio, Fib, TT and PLT showed no significant difference between effective chemotherapygroup and non-effective chemotherapy group.3. Clinical research on coagulation index used to evaluate the tumor metastasis riskthrough blood circulationMethod: The15ml venous blood was drawn in90female patients with pathologicallydiagnosed stage Ⅰ～Ⅲ breast cancer and17stage â…£ breast cancer patients undergoing no treatment. PBMCs （peripheral blood monouclear cells） were gathered from the blood bydensity gradient centrifugation, and then CTCs with CK （Cytokeratin）19mRNA positive inPBMCs were detected by RT-PCR. Meanwhile the blood coagulation index was detected inall these patients. After this, the discrepancy of blood coagulation index between CTCnegative group and CTC positive group, and CTC detection rate were contrastively analyzed.Results:1) Fib level was （2.67±0.44） g/L in CTC negative group,（3.03±0.81） g/Lin CTC positive group; there was a significant difference between the two groups （P=0.004）.PLT was （185.15±46.22）×10⁹/L in CTC negative group,（205.08±51.16）×10⁹/L in CTCpositive group; there was a significant difference between the two groups （P=0.041）. PT was（10.71±0.57） s in CTC negative group,（11.01±0.57） s in CTC positive group; there was asignificant difference （P=0.011） between the two groups. PT%was108.37±12.81in CTCnegative group,101.44±13.01in CTC-positive group; there was a significant difference （P=0.009） between the two groups. PTINR was0.93±0.05in CTC negative group,0.96±0.05in CTC positive group; there was a significant difference between the two groups （P=0.005）.APTT, APTT Ratio, DD and TT showed no statistical difference between CTC negativegroup and CTC-positive group （P>0.05）.2)101patients presented Fib≤3.70g/L, including36patients with CTC positive （35.64%）; six patients presented Fib>3.70g/L, including5patients of CTC positive （83.33%）; there was a significant difference between the two groups（P=0.020）. CTC detection rate showed no statistical difference by grouping based on PT,PT%, PTINR, APTT, APTT Ratio, DD, TT, PLT （all P values>0.05）.Conclusion1. Compared with the patients with breast benign disease, breast cancer patients werewith shorter PTINR,higher Fib and D-D level,longer TT; Compared with the patients withbreast cancer of stage I-III, the patients in stage â…£ were more likely to have a higher bloodFib level; The Fib level of group T4was higher than that of group T₁-T₃; The Fib level ofgroup N3was higher than that of group N₀-N₂; The patients with HER-2positive had longerAPTT and TT, and higher D-D level.2. The Fib and PLT increased while the D-D decreased after chemotherapy; The D-Dlevel at baseline in effective chemotherapy group was higher than that in non-effective group;the patients with D-D decreased after chemotherapy presented higher remission rate. 3. Compared with CTC negative group, in CTC positive group, Fib and PLT levels wereincreased, PT increased, PT%decreased and PTINR increased. CTC positive ratio in patientswith Fib＞3.70g/L was higher than that of Fib≤3.70g/L group.