Secretomic profiling of cells from hollow fiber bioreactor reveals PSMA3 as a potential cholangiocarcinoma biomarker

Abstract

Cholangiocarcinoma (CCA), derived from the bile duct, occurs with a relatively high incidence in Northeast Thailand. Early diagnosis is still hampered by the lack of sufficient biomarkers. In recent years, biomarker discovery using secretomes has provided interesting results, including our studies on CCA secretomes, especially with three-dimensional cell cultures. Thus, cells cultured using the hollow fiber bioreactor (HFB) with 20 kDa molecular weight cut-off (MWCO) yielded higher quality and quantity of secretomes than those from conditioned media of the monolayer culture (MNC) system. In this study, we employed the HFB culture system with 5 kDa MWCO and compared conditioned media from the HFB and MNC systems using two-dimensional gel electrophoresis, followed by identifying proteins of interest by liquid chromatography and mass spectrometry (LC/MS/MS). Two out of 4 spots of NGAL or lipocalin-2 were found to show highest increase in expression of 19.93-fold and 18.79-fold in HFB compared to MNC. Interestingly, all 14 proteasome subunits including proteasome subunit α type-1 to type-7 and β type-1 to type-7 showed 2.92-fold to 12.13-fold increased expression in HFB. The protein-protein interactions of upregulated proteins were predicted, and one of the main interaction clusters involved 20S proteasome subunits. Proteasome activity in the HFB conditioned media was also found to be higher than that in MNC conditioned media. Three types of proteasome subunit were also validated by immunoblotting and showed higher expression in the HFB system compared to MNC system. Proteasome subunit α type-3 (PSMA3) showed the highest level in plasma of cholangiocarcinoma patients compared to normal and hepatocellular carcinoma patients by immunodetection, and is of interest as a potential biomarker for cholangiocarcinoma.

Prognostic and Genomic Analysis of Proteasome 20S Subunit Alpha (PSMA) Family Members in Breast Cancer

“…PSMA3 , alternatively known as proteasome subunit alpha type 3, encodes the α7 subunit, which makes up the outer ring of the 20S core particle. Cancer-related reports on the relation of PSMA3 expression and malignancy are limited, including metastatic gastric cancer and cholangiocarcinoma [ 82 , 83 ]. PSMA4 , alternatively known as proteasome subunit alpha type 4, encodes the α3 subunit, which makes up the outer ring of the 20S core particle.…”

MLN2238 exerts its anti-tumor effects via regulating ROS/JNK/mitochondrial signaling pathways in intrahepatic cholangiocarcinoma

“…Proteasome-related genes play key roles in the formation of proteasomal structures and in maintaining its activity. Recent studies have revealed that some proteasome-related genes, such as PSMA3 and PSMC2, can promote the progression of malignant biliary tumors and act as potential biomarkers to predict prognosis ( Verathamjamras et al, 2017 ; Duan et al, 2021 ; Zhu et al, 2021 ). To fully explore the function of the proteasome in the progression of iCCA, 43 proteasome-related genes were included, and the hierarchical heatmap showed that these genes, such as PSMD1, PSMB4, PSMD13, and PSMC4, were out of balance during trajectory development ( Figure 2G ).…”

Immuno‐oncological role of 20S proteasome alpha‐subunit 3 in aggravating the progression of esophageal squamous cell carcinoma

“…Furthermore, several studies have demonstrated the abnormal expression profiles of PSMA3 in several cancers and identified PSMA3 as a cancer-related indicator [11]. An analysis of plasma samples from normal individuals, cholangiocarcinoma (CCA) patients, and hepatocellular carcinoma patients was conducted by Verathamjamras et al [12], and the results provided evidence that PSMA3 could be a potential biomarker for CCA. Also, Ding et al reported that PSMA3 was explicitly enriched in the serum of patients with metastatic gastric cancer [13].…”

“…An analysis of plasma samples from normal individuals, cholangiocarcinoma (CCA) patients, and hepatocellular carcinoma patients was conducted by Verathamjamras et al. [12], and the results provided evidence that PSMA3 could be a potential biomarker for CCA. Also, Ding et al.…”