Integrative analysis revealed a correlation of PIAS family genes expression with prognosis, immunomodulation and chemotherapy.

BACKGROUND: Protein inhibitor of activated STATs (PIAS) has pleiotropic biological effects, such as protein post-translational modification, transcriptional coregulation and gene editing. It is reported that PIAS family genes are also correlated with immune cells infiltration in cancers that highlights their unnoticed biological role in tumor progression. However, the relationship of their expression with prognosis, immune cell infiltration, tumor microenvironment, and immunotherapy in pan-cancer has been rarely reported. METHODS: The multi-omics data were used to investigate the expression level of PIAS family members in pan-cancer, and the prognostic value of their expression in different tumors was analyzed by univariate Cox regression and Kaplan-Meier. Correlation analysis was used to investigate the relationship of PIAS gene expression with tumor microenvironment, immune infiltrating subtypes, stemness score and drug sensitivity. In addition, we also used wound healing and transwell assays to verify the biological effects of PIAS family gene expression on invasion and metastasis of HCC cells. RESULTS: We found that PIAS family genes expression is significantly heterogeneous in tumors by multi-genomic analysis, and associated with poor prognosis in patients with multiple types of cancer. Furthermore, we also found that genetic alterations of PIAS family genes were not only common in different types of human tumors, but were also significantly associated with disease-free survival (DFS) across pan-cancer. Single-cell analysis revealed that PIAS family genes were mainly distributed in monocytes/macrophages. Additionally, we also found that their expression was associated with tumor microenvironment (including stromal cells and immune cells) and stemness score (DNAss and RNAss). Drug sensitivity analysis showed that PIAS family genes were able to predict the response to chemotherapy and immunotherapy. PIAS family genes expression is closely related to tumor metastasis, especially PIAS3. High PIAS3 expression significantly promotes the migration and invasion of liver cancer cell lines (HCC-LM3 and MHCC97-H). CONCLUSIONS: Taking together, these findings contribute to determine whether the PIAS family genes are a potential oncogenic target gene, which have important contribution for the development of cancer immunotherapy.

[Research on Ex Vivo Hematopoiesis Supported by Microcarriers]

To explore methods of maintaining the self-renewal capacity of hematopoietic stem cells, inhibiting their overdue differentiation and expanding hematopoietic cells massively, the murine bone marrow stromal cells were coated on microcarriers, then co-cultured with hematopoietic cells from murine bone marrow as group 2 (G2). The G2 contents were wrapped by sodium alginate, then cultivated as group 1 (G1). The only microcarriers coated with stromal cells as group 3 (G3) and the only bone marrow cells as group 4 (G4) were cultivated as control groups. Contrasting observation and microphotograph were performed; the number of total marrow cells, the colony efficiency of CFU-GM and the percentages of CD34(+) cells were determined. Three repeated experiments indicated that the colony efficiency of CFU-GM before culture (G0) were 118.8 +/- 38.1/10(5) marrow cells, and the total outputs of CFU-GM (G0) were 9 501.3 +/- 3 049.0. After culture for two weeks, hematopoietic cells were adhered to or embedded in stromal cells coating the microcarriers, and had formed hematopoietic focus. The colony efficiency of CFU-GM per 10(5) mononuclear cells in group G1, G2, G3 and G4 averaged 30.9 +/- 13.7, 147.3 +/- 66.0, 23.4 +/- 23.1 and 15.9 +/- 8.1, respectively; the total outputs of CFU-GM in group G1, G2, G3 and G4 averaged 273.8 +/- 75.3, 9 424.8 +/- 7 933.7, 419.1 +/- 305.6 and 140.7 +/- 20.7, respectively; the measured CFU-GM output in group G2 was significantly higher than that in group G4, and still significantly higher than the sum of groups G3 and G4 (t = 6.553, t = 5.494; P < 0.05). The percentage of CD34 cells before culture was 10.0 +/- 1.0; after cultuer for two weeks, the percentages of CD34(+) cells in G1, G2, G3 and G4 averaged 4.0 +/- 1.0, 11.0 +/- 1.0, 3.3 +/- 1.5 and 2.2 +/- 0.8, respectively. The percentage of CD34 positive control (3T3 cells) was 17.0 +/- 1.0. This result was consistent with the result of CFU-GM outputs measured. These data suggest that microcarriers coated with stromal cells can perfectly support the ex vivo hematopoiesis at least to four weeks, while hematopoietic cells fixed by alginate are not significantly different from control groups. The hematopoiesis-simulating model of microcarriers is successful, whereas the hematopoiesis-simulating model of alginate macrocarriers can not support the ex vivo hematopoiesis.