Hepatocellular carcinoma (HCC) is the most common form of primary liver cancer. γδ T cells have been revealed to be promising candidates for immunotherapy in patients with HCC. However, the use of these cells in clinical practice has been demonstrated to be challenging. In the present study, γδ T cells isolated from the peripheral blood of patients with HCC (n=83) and healthy donors (n=15) were characterized. Flow cytometry was used to analyze the proportion, phenotype, tumor-killing capacity and cytokine secretion of regulatory T cells (Tregs) and γδ T17 cells in peripheral blood samples prior to and following amplification. Interleukin (IL)-17A levels in the supernatant was analyzed using an ELISA on days 3, 7, 10 and 14. The in vitro cytotoxicity of γδ T cells was measured using an MTT assay. It was revealed that zoledronate with IL-2 may efficiently expand γδ T cells sourced from the peripheral blood of patients with HCC. The amplification capacity of γδ T cells was associated with the clinicopathological characteristics of patients (clinical stage, levels of AFP and albumin, duration of disease, size and number of tumors, numbers of Tregs and γδ T17 cells, and levels of IL-17A). The proportion of γδ T cells positive for interferon-γ, tumor necrosis factor-α, granzyme B, perforin, and lysosome-associated membrane protein 1 was almost unchanged prior to and following amplification. Following amplification, the in vitro cytotoxicity of γδ T cells also remained unchanged. γδ T17 cells, Tregs and IL-17A levels were not altered during amplification. In summary, following in vitro amplification, circulating γδ T cells were revealed to possess features that may make them suitable for immunotherapy for HCC without increasing immunosuppressive factors. However, immunotherapy should be individualized according to the clinicopathological features of patients.

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A major obstacle to effective cancer immunotherapy is the tumor immune microenvironment. Natural killer (NK) cell resistance has been suggested as a primary cause of poor prognosis in hepatocellular carcinoma (HCC), which seemingly correlates with CNOT7 overexpression. CNOT7, a cytoplasmic mRNA deadenylase that is highly expressed in HCC, may regulate cytokine transforming growth factor‐β1 (TGF‐β1) secretion by controlling nuclear factor‐κB subunit p65 trafficking. CNOT7 depletion suppresses TGF‐β1 secretion in HCC and promotes interferon‐γ (IFN‐γ) secretion by NK cells, and we previously demonstrated that CNOT7 depletion reversed IFN‐γ resistance in HCC cells. Therefore, we hypothesized that CNOT7 depletion might reverse NK cell resistance by influencing the tumor immune microenvironment of HCC. To test this hypothesis, we examined the correlation between CNOT7, STAT1, TGF‐β1 and IFN‐γ expression with hepatitis B virus‐related cirrhosis and HCC with hepatitis B virus‐related cirrhosis. We found that modulation of CNOT7 expression alters TGF‐β1 secretion in HCC and IFN‐γ secretion in NK cells. We also examined the effects of NK cells in HepG2 cells with CNOT7 knockdown, which showed that NK cell surface CD107a expression is up‐regulated and caspase‐3 expression is significantly enhanced in CNOT7‐deficient HepG2 cells. Overall, our results show that knockdown of CNOT7 expression reverses NK cell resistance in HCC cells. Therefore, CNOT7 depletion has potential as a new adjuvant therapy in immunotherapy for HCC.

Colorectal cancer (CRC) is the third cause of cancer-related death and the fourth most frequently diagnosed cancer across the globe. The objective of this study is to obtain novel and effective diagnostic markers to enrich CRC diagnosis methods. Herein, exosomal miRNA expression data of CRC and normal blood were subjected to XGBoost algorithm, and 5 miRNAs related to CRC diagnosis were primarily confirmed. Then multilayer perceptron (MLP) classifiers were constructed based on different subsets. Via integrated feature selection (IFS), we noticed that the MLP classifier constructed by the first four miRNAs (miR-654-5p, miR-126, miR-10b, and miR-144) had the highest Matthews correlation coefficient (MCC). Subsequently, principal component analysis (PCA) for dimensionality reduction was performed on samples based on the miR-654-5p, miR-126, miR-10b, and miR-144 expression data. The signature based on these four feature miRNAs, as the analysis indicated, could effectively distinguish CRC samples from normal samples. Further, we extracted the exosomes from clinical blood samples and applied qRT-PCR analysis, which revealed that the expression of these four feature miRNAs was in the trend of that in the test set. Collectively, these four feature miRNAs might be tumor biomarkers in the serum, and our study offers innovative thinking on early-stage CRC diagnosis.