Enhanced procoagulant activity of platelets after chemotherapy in non-small cell lung cancer

Figures & data

Table 1. Characters of patients with NSCLC before chemotherapy.

	Control (n = 52)	Patients (n = 52)
Age (years)	51 ± 7	66 ± 5
Sex (Male/Female)	28/24	31/21
Temperature (°C)	36.3 ± 0.4	36.7 ± 0.7
Respiratory Rate (bpm)	14 ± 2	23 ± 6
Heart Rate (bpm)	79 ± 12	74 ± 24
NSCLC histology
Adenocarcinoma	—	27
Squamous	—	14
Large cell	—	2
NSCLC NOS	—	9
Clinical Stage
IA / B	—	22
IIA / B	—	24
IIIA/ B	—	5
IV	—	1
RBC (10^12/L)	4.2 ± 1.6	2.7 ± 2.1
Hb (g/L)	142 ± 36	103 ± 22^*
WBC (10^9/L)	3.4 ± 2.1	7.2 ± 3.5
Platelet (10^9/L)	215 ± 56	307 ± 89^*
PT (s)	12.8 ± 2.7	14.8 ± 4.4
aPTT (s)	23.5 ± 3.6	15.2 ± 5.8^*
TAT (ng/ml)	142.6 ± 27	537 ± 31.5^*
Fibrinogen (g/L)	3.1 ± 1.2	3.7 ± 4.1
D-dimer (g/L)	0.9 ± 0.7	6.6 ± 2.7^*

Values given in mean and SD.

* P < 0.05 vs. control. Abbreviations: RBC, red blood cell; Hb, hemoglobin; WBC, white blood cell; PT, prothrombin time; APTT, activated partial thromboplastin time.

Table 2. Characters of platelets in healthy controls and NSCLC patients before chemotherapy.

	Control	Patients
PS expression (%)	1.5 ± 0.6	4.9 ± 2.3^*
PMP (/μl)	1652 ± 177	3216 ± 325^**
Coagulation time (s)	315 ± 23	247 ± 31^*
Thrombin (nM)	1.7 ± 0.9	5.8 ± 2.1^**

Values given in mean and SD.

* P < 0.05,

** P < 0.01 vs. control. Abbreviations: PS, phosphatidylserine; PMP, platelet microparticle.

Figure 1. Clotting time and TAT in patients with NSCLC within 7 d after chemotherapy. Blood of patients before and after chemotherapy was isolated, and clotting time of whole blood (a) and TAT level in serum (b) were measured. Data are present as mean ± SD, *P < 0.05 vs day 0, **P < 0.01 vs day 0.

Figure 2. PS expression and PCA of platelets after chemotherapy. Platelets were isolated from patients before and after chemotherapy. (a) PS and TF expression on platelets were measured respectively with Alexa 488-lactadherin (green) and Alexa 647-anti CD142 (red) by flow cytometry. (b) Coagulation time was measured by coagulometer and thrombin and fibrin formation were evaluated with microplate reader. Data are present as mean ± SD, *P < 0.05 vs day 0, **P < 0.01 vs day 0.

Figure 3. The number of PMPs after chemotherapy. Platelets were isolated from patients before and after chemotherapy. (a,b) The number of total MPs and PMPs were analyzed by flow cytometry. PMPs were characterized by CD41a expression. Data are present as mean ± SD, **P < 0.01 vs day 0. (c) Confocal microscopy images showed quiescent platelets (left, arrow) and apoptotic platelets positive for lactadherin staining (right, arrow) with MPs (right, arrowheads) at day 1. Bars represent 2 µm.

Figure 4. The effect of cisplatin on PCA of platelets. Platelets were isolated from healthy subjects and treated with cisplatin of different concentrations for 1 h in vitro. Coagulation time (a), intrinsic/extrinsic FXa and thrombin (b) as well as fibrin (c) formation were evaluated. Data are present as mean ± SD, *P < 0.05 vs untreated, **P < 0.01 vs untreated. (c) After 1µM cisplatin treatment of 1 h, quiescent platelets (left), activated platelets (middle) and apoptotic platelets with MPs (right) were shown by scanning microscopy images. Bars represent 1 µm.

Figure 5. The effect of anti-platelet agents and lactadherin on the PCA. After 1 µM cisplatin treatment of 1 h, platelets were treated with aspirin (100 nM) and lactadherin (2 nM). Coagulation time (a), intrinsic/extrinsic FXa and thrombin (b) as well as fibrin (c) formation of platelets were measured. Data are present as mean ± SD, *P < 0.05 vs control, **P < 0.01 vs control.