Positive effect of RORγt on the prognosis of thyroid papillary carcinoma patients combined with Hashimoto's thyroiditis

Abstract

Objectives: To investigate the positive effect of Th17 cells on the prognosis of patients with PTC and HT.

Methods: The expression of nuclear specific marker RORγt of Th17 cells in fresh and paraffin thyroid tissues and serum specimens were analyzed. Flow cytometry was used to detect the formation rates of Th17 cells (CD3⁺CD8^-IL-17A⁺/CD3⁺CD8^-%) at different time points after co-culture of thyroid papillary carcinoma cell line (TPC-1 and K1) and umbilical cord blood initial T lymphocytes. The protein expression of RORγt in T lymphocytes after co-culture was detected. Preoperative serum levels of Th17 (IL-17) cytokines were measured.

Results: The positive expression of RORγt in the tumor microenvironment of PTC patients with or without HT could inhibit the lymph node metastasis of the tumor. PTC cancer cells could induce initial T lymphocyte to differentiate into Th17 cells, and the K1 cell line with lymph node metastasis induced a higher proportion of RORγt protein than that in TPC-1 cell line without lymph node metastasis. In PTC patients with HT, serum IL-17 concentration was negatively correlated with lymph node metastasis in the central group.

Conclusions: RORγt may play an anti-tumor role in reducing thyroid cell damage by reducing the thyroid autoimmune antibodies TPOAb and TGAb in the PTC population in Yunnan plateau region.

Keywords: Hashimoto thyroiditis; Papillary thyroid carcinoma; prognosis.

Figure 1

Schematic diagram of transwell chamber isolation co-culture.

Figure 2

Th17 (CD3⁺CD8^-IL-17⁺) lymphocyte in umbilical cord blood flow cytometry gate (A: CD45⁺ lymphocyte in umbilical cord blood; B: CD3⁺ lymphocyte; C: CD3⁺CD8^-IL-17⁺ lymphocyte; D: Th17 (CD3⁺CD8^-IL-17⁺) lymphocyte).

Figure 3

Expression of RORγt in thyroid papillary carcinoma with or without Hashimoto’s thyroiditis microenvironment (A: PTC alone ×100; B: PTC alone ×400; C: PTC with HT ×100; D: PTC with HT ×400).

Figure 4

mRNA expressive abundance of RORγt gene in cancer tissue associated with PTC and PTC/HT patients (NS: No statistical significance).

Figure 5

The percentage of Th17 cells induced by co-culture for 24 h, 36 h, 48 h and 60 h after isolation with Umbilical cord blood initial T lymphocytes and papillary thyroid carcinoma cell lines (A: TPC-1 and B: K1), normal thyroid follicular epithelial cell lines (C: Nthy ori 3-1).

Figure 6

Flow cytometry detection of the proportion change trend of inductive formation CD3⁺CD8^-IL-17⁺Th17 cells after isolation co culture of cord blood initial T cells combined with papillary thyroid cancer cell lineage (TPC-1 and K1) and normal thyroid follicular epithelial cell line (Nthy ori 3-1) treated for 24 h, 36 h, 48 h, 60 h. The results of co-culture with Nthy ori 3-1 were 7.26 ± 2.14 in 0 h, 9.07 ± 1.33 in 24 h, 10.70 ± 1.45 in 36 h, 8.70 ± 1.87 in 48 h, 9.57 ± 1.29 in 60 h. The results of co-culture with TPC-1 were 7.85 ± 1.50 in 0 h, 18.77 ± 1.45 in 24 h, 24.00 ± 1.68 in 36 h, 13.83 ± 2.25 in 48 h, 9.57 ± 1.29 in 60 h. The results of co-culture with K1 were 7.88 ± 2.84 in 0 h, 19.63 ± 1.81 in 24 h, 43.30 ± 2.86 in 36 h, 24.03 ± 1.78 in 48 h, 32.63 ± 3.25 in 60 h. Symbol *indicate significance at the 0.05 level, symbol **indicate significance at the 0.01 level, symbol ***indicate significance at the 0.001 level.

Figure 7

Flow cytometry detection of the proportion change trend of inductive formation Th17 cells after isolation co culture of cord blood initial T cells combined with papillary thyroid cancer cell lineage (TPC-1 and K1) and normal thyroid follicular epithelial cell line (Nthy ori 3-1) treated for 24 h, 36 h, 48 h, 60 h.

Figure 8

Expression level of IL-17 in peripheral blood serum of patients in benign tumor control (BTD) group 11.28 ± 10.86 pg/ml, Hashimoto’s thyroiditis (HT) group 22.45 ± 12.47 pg/ml, thyroid papillary carcinoma with Hashimoto’s Thyroiditis (PTC/HT) group 15.16 ± 10.61 pg/ml and papillary thyroid carcinoma (PTC) group 12.64 ± 11.07 pg/ml.

Figure 9

Dynamic-Gene Net of differential Immune-related gene (RORγt gene was positively correlated with CAV1, the interaction coefficient = 0.93. The interaction coefficient is the degree of co-expression of two genes).