JAK2 inhibitor combined with DC-activated AFP-specific T-cells enhances antitumor function in a Fas/FasL signal-independent pathway.

OBJECTIVE: Combination therapy for cancer is more effective than using only standard chemo- or radiotherapy. Our previous results showed that dendritic cell-activated α-fetoprotein (AFP)-specific T-cells inhibit tumor in vitro and in vivo. In this study, we focused on antitumor function of CD8(+) T-cells combined with or without JAK2 inhibitor. METHODS: Proliferation and cell cycle were analyzed by CCK-8 and flow cytometry. Western blot was used to analyze the expression level of related protein and signaling pathway. RESULTS: We demonstrated reduced viability and induction of apoptosis of tumor cells with combination treatment. Intriguingly, cell cycle was blocked at the G1 phase by using AFP-specific CD8(+) T-cells combined with JAK2 inhibitor (AG490). Furthermore, an enhanced expression of BAX but no influence on Fas/FasL was detected from the tumor cells. CONCLUSION: These results indicate a Fas/FasL-independent pathway for cellular apoptosis in cancer therapies with the treatment of AFP-specific CD8(+) T-cells combined with JAK2 inhibitor.

Circulating Hepatocellular Cells are a Bad Prognostic Factor for HCC Patients.

BACKGROUND/AIMS: Circulating hepatocellular carcinoma cells (CHCCs) may be detected by reverse transcription-polymerase chain reaction (RT-PCR). We investigated the relationship between CHCCs and hepatoma patients' survival period after different managements. METHODOLOGY: Peripheral blood (5 ml) samples were obtained from 93 patients with hepatocellular carcinoma (HCC), and from 33 control subjects (9 with liver cirrhosis after hepatitis B, 14 with chronic hepatitis B, 10 with healthy people) between January 1st, 2009 and December 31, 2012. To detect CHCCs in peripheral blood, alpha-fetoprotein (AFP) messenger RNA (mRNA) was amplified from total RNA extracted from whole blood by RT-PCR. RESULTS: AFPmRNA was detected in 49 blood samples from the HCC patients (49/93, 53.0%). In contrast, there were no clinical control subjects whose samples showed detectable AFPmRNA. The presence of AFPmRNA in blood seemed to be correlated with the tumor stage (by TNM classification) of HCC, the serum AFP value, and the presence of intrahepatic metastasis, portal vein thrombosis, tumor diameter and/or distant metastasis. CONCLUSIONS: The presence of AFP mRNA in peripheral blood may be an indicator of CHCCs, which might predict hematogenous spreading metastasis in patients with HCC and may be as a bad prognostic factor for HCC patients.

Significance of detecting circulating hepatocellular carcinoma cells in peripheral blood of hepatocellular carcinoma patients by nested reverse transcription-polymerase chain reaction and its clinical value: a retrospective study.

AIMS AND BACKGROUND: Circulating hepatocellular carcinoma cells may be detected by reverse transcription-polymerase chain reaction. We investigated the relationship between circulating hepatocellular carcinoma cells and hepatoma patient survival after different managements and survival periods. METHODS AND STUDY DESIGN: Peripheral vein blood (5 ml) samples were obtained from 113 patients with hepatocellular carcinoma and from 33 control subjects (9 with liver cirrhosis after hepatitis B, 14 with chronic hepatitis B, 10 healthy individuals) between January 1, 2009, and December 31, 2013. To detect circulating hepatocellular carcinoma cells in peripheral blood, alpha-fetoprotein messenger RNA was amplified from total RNA extracted from whole blood by reverse transcription-polymerase chain reaction. RESULTS: Alpha-fetoprotein messenger RNA was detected in 59 blood samples from the hepatocellular carcinoma patients (59/113, 52.2%). In contrast, there were no clinical control subjects whose samples showed detectable alpha-fetoprotein messenger RNA. The presence of alpha-fetoprotein messenger RNA in blood seemed to be correlated with the stage (by TNM classification) of hepatocellular carcinoma, serum alpha-fetoprotein value, and the presence of intrahepatic metastasis, portal vein thrombosis, tumor diameter and/or distant metastasis. In addition, alpha-fetoprotein messenger RNA was detected in the blood of 25 patients showing distant metastasis at extrahepatic organs (100%), in contrast to 32 of 88 cases without metastasis (36.4%). All the patients with hepatocellular carcinoma were followed. Seventeen patients with resection of a T 2 stage hepatocellular carcinoma had a survival of 3.2 years after surgical management, 38 cases with resection of a T3 stage hepatocellular carcinoma had a 1.3-year survival, and only 37 cases with T4 stage disease after different treatments except surgery survived for 0.6 years (P <0.01). CONCLUSIONS: The presence of alpha-fetoprotein messenger RNA in peripheral blood may be an indicator of circulating hepatocellular carcinoma cells, which might predict hematogenous spreading metastasis in patients with hepatocellular carcinoma and may be a poor prognostic factor for hepatocellular carcinoma patients.