I-Simultaneous evolution of the proteasome in serum according to the chymotrypsin-like activity

Serum Proteasome measured by ELISA, in 145 patients (with an average age 47.9 ± 15.3 years) suffering from Multiple Myeloma,Lymphoma, Acute Leukemya and Chronic Leukemia according to the chymotrypsin-like activity is represented in Fig. 1.

Fig. 1

Correlation between serum concentration of proteasome and the chymotrypsin-like activity: in patients with AL1 and CL2, LYMPHOMA3, andMM4

All patients with Hematologic malignancies express proteolysis rate more pronounced in serum compared to the control.

However, analysis of the correlation between chymotrypsin-like activity and the concentration of the serum proteasome shows no logical link with all types of Hematologic Diseases.

II-Simultaneous evolution of the subcellular proteasome according to the chymotrypsin-like activity

The rate of intracellular Proteasome, in patients, reached to hematological malignancies according to the chymotrypsin-like activity is represented in Fig. 2

Fig. 2

Correlation between intracellular concentration of proteasome and the chymotrypsin-like activity: in patients with AL1 and CL2, LYMPHOMA3, andMM4

Furthermore, the analysis of changes in intracellular chymotrypsin-like activity based on intracellular proteasome (ng/ml), reveals a positive evolutionary trend and those in all recruited patients (CL, AL, MM and Lymphoma).

III-Influence of treatment on the evolution of the proteasome at the Hematological malignanciesProteasome concentration and chymotrypsine-like activity in serum

The proteasome, a key element of the neoplastic differentiation, currently attracting great interest as a new marker in different pathologies.

A population of 60 patients with different forms of Hematologic Diseases was recalled for a second blood test after three months of treatment.

After 3 months of treatment, all patients with Hematological malignancies, express a serum proteasome levels significantly low compared to stage I (no treatment).The patients with MM and AL show a very significant regression respectively 57% and 45%. A reduction in serum catalytic activity was observed in all patients who have undergone treatment. A sharp drop in catalytic activity (60%) was observed in patients with AL. The patients with lymphoma, MM, CL, display a respective decrease of 47%, 37%, 35% (Fig. 3).

Fig. 3

Change in serum concentration of proteasome and proteolytic activity depending on the type of pathology (I: before treatment; II: under treatment)

Subcellular rate of proteasome and chymotrypsine-like activity

All Subjects with Hematologic malignancies, having received an anti-tumor treatment, recalled after a three-month period, have levels of intracellular proteasome significantly reduced compared to the values of stage I (untreated).

In fact there was a decrease in intracellular concentration of the most important proteasome in patients with CL, about 41%, followed by lymphoma patients display decreased proteasome rate of 29%, followed by the patients with AL showing a decrease of 16.5%; and patients with myeloma, which has the lowest rate reduced proteasome 10%

we find that patients with CL, shows a highest percentage decrease in catalytic activity (about 46%) after taking cancer treatment, followed those with Myeloma display a decrease in the catalytic activity of about 40%, patients with AL with a reduction rate of 31% and lymphoma patients with a decrease of 25% of the catalytic activity.(Fig. 4).

Fig. 4

Change in sub cellular concentration of proteasome and proteolytic activity depending on the type of pathology (I: before treatment; II: under treatment)

Quantitative and functional evolution of the proteasome depending on the origin of tumor cells

We subdivided our cancer patient population into two categories based on the origin of the tumor cell line: myeloid lineage (n = 54) included patients with AML and CML and Lymphoid line (n = 92) Lymphoma, Myeloma and CLL.A significant decrease in serum levels of the proteasome was observed in the Myeloid lineage (concentration: 4116 ± 702 ng/ml) compared to the Lymphoid lineage (concentration: 5554 ± 512 ng/ml; p < 0.05).

In addition, there was a significant difference (P < 0.001) in the mean intracellular proteolytic activity between the Lymphoid and Myeloid line.

No significant difference (p > 0.05) was observed between the Lymphoid and Myeloid line in the subcellular proteasome concentration or serum proteolytic activity (Fig. 5).

Fig. 5

Comparison of the rate and catalytic activity of proteasome between Myeloid and Lymphoid Lineage at sub cellular and serum level; *: P < 0.05 compared to the myeloid lineage

Quantitative and functional evolution of the proteasome depending on the evolutionary stage of the disease in lymphomaafter-treatment

We classified our patient population suffering from lymphoma (n = 22), depending on the stage of tumor progression (according to Ann Arbor classification) intotwo types (1: Class I and II, and 2: Class III).

After that we analyzed the quantitative and functional evolutionary trend of the proteasome after taking the treatment, we noted that patients suffering from lymphoma class III report a significant regression (49%) after treatment (before: 6450 ± 940 ng/ml; after 3600 ± 560 ng/ml in the patients with NHL;before 6500 ± 760 ng/ml; after 3850 ± 550 ng/ml in patients with HL).Otherwise a regression(27%) is recorded in stage I and II lymphoma (NHL and HL). The same situation is observed at the chymotrypsin-like activity (Fig. 6).

Fig. 6

Comparison of the rate and catalytic activity of proteasome between LNH and LH in sub cellular level

Functional evolution of proteasome according to a biological parameters (LDH)

In patients with hematological malignancies, a biological parameters was tested and analyzed, in order to establish possible statistical correlation with the level of circulating and intracellular 20 s proteasome: Lactate Dehydrogenase (LDH) as global marker of cytolysis.

A progressive evolutionary trend is observed between the activity of the proteasome and LDH released into circulation (Fig. 7). Otherwise the linear correlation report significant dependence average (R2 = 0.699).

Fig. 7

Simultaneous Evolution of serum chymotrypsin-like activity (UF) and the rate of LDH (UI/ l) in patients with Hematologic malignancies