Muhanna Nidal, Amer Johnny, Salhab Ahmad, Sichel Jean-Yves, Safadi Rifaat
Liver and Gastroenterology Units, Division of Medicine, Hadassah University Medical Center, Jerusalem, Israel; Department of Otolaryngology, Head & Neck Surgery, Shaare Zedek Medical Center, Jerusalem, Israel.
Liver and Gastroenterology Units, Division of Medicine, Hadassah University Medical Center, Jerusalem, Israel.
PLoS One. 2015 Jul 7;10(7):e0132463. doi: 10.1371/journal.pone.0132463. eCollection 2015.
A high prevalence of thyroid papillary cancer was reported in hepatitis-C-virus (HCV) positive patients. However, the mechanistic role of hepatic-fibrosis in thyroid malignancy progressions is still unclear.
We aimed to study the immune-modulatory interactions between thyroid papillary carcinoma and hepatic-fibrosis.
Hepatic-fibrosis was induced in nude-nu-male mice by intra-peritoneal administration of carbon-tetrachloride. To induce thyroid-tumor, a thyroid papillary carcinoma cell line (NPA) was injected subcutaneously in the backs. Fibrotic profile was estimated by α-smooth-muscle-actin (αSMA) expression in liver tissue extracts using western-blots and RT-PCR. Intra-hepatic NK cells were isolated and stained for NK activity (CD107a) by flow cytometry. Liver histopathology (H&E staining), thyroid tumor mass and serum alanine aminotransferase (ALT), serum vascular endothelial growth factor (VEGF) and free-T4 levels were also assessed.
Ex-vivo: NPA cells were co-cultured with intra-hepatic NK cells isolated from fibrotic mice with/without the tumor were analyzed for CFSE-proliferations. Both tumor groups (with/without hepatic-fibrosis) excreted higher serum free T4 levels. Hepatic-fibrosis increased tumor weight and size and serum free-T4 levels. In addition, tumor induction increased liver injury (both hepatic-fibrosis, necro-inflammation and serum ALT levels). In addition, tumor-bearing animals with hepatic-fibrosis had increased NK activity. NPA tumor-bearing animals increased fibrosis in spite of increased NK activity; probably due to a direct effect through increased serum free-T4 excretions. Serum VEGF levels were significantly increased in the fibrotic- bearing tumor groups compared to the non-fibrotic groups. In-vitro, NK cells from fibrotic tumor-bearing animals reduced proliferation of NPA cells. This decrease is attributed to increase NK cells activity in the fibrotic animals with the NPA tumors.
Our results propose that NK cells although were stimulated in advanced fibrosis with tumor, they lost their anti-tumor and anti-fibrotic activity probably due to secretions of T4 and VEFG and may explain increased risk of thyroid tumors in chronic HCV patients.
据报道，丙型肝炎病毒（HCV）阳性患者中甲状腺乳头状癌的患病率较高。然而，肝纤维化在甲状腺恶性肿瘤进展中的作用机制仍不清楚。
我们旨在研究甲状腺乳头状癌与肝纤维化之间的免疫调节相互作用。
通过腹腔注射四氯化碳在裸鼠雄性小鼠中诱导肝纤维化。为了诱导甲状腺肿瘤，将甲状腺乳头状癌细胞系（NPA）皮下注射到背部。使用蛋白质免疫印迹法和逆转录-聚合酶链反应通过肝组织提取物中的α-平滑肌肌动蛋白（αSMA）表达来评估纤维化情况。分离肝内自然杀伤细胞（NK细胞），并通过流式细胞术对NK活性（CD107a）进行染色。还评估了肝脏组织病理学（苏木精-伊红染色）、甲状腺肿瘤大小以及血清丙氨酸氨基转移酶（ALT）、血清血管内皮生长因子（VEGF）和游离T4水平。
体外实验：将NPA细胞与从患有或未患有肿瘤的纤维化小鼠中分离的肝内NK细胞共培养，分析其羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）增殖情况。两个肿瘤组（有/无肝纤维化）的血清游离T4水平均较高。肝纤维化增加了肿瘤重量、大小和血清游离T4水平。此外，肿瘤诱导增加了肝损伤（肝纤维化、坏死性炎症和血清ALT水平）。此外，患有肝纤维化的荷瘤动物的NK活性增加。尽管NK活性增加，但携带NPA肿瘤的动物的纤维化仍增加；这可能是由于血清游离T4排泄增加的直接作用。与非纤维化组相比，纤维化荷瘤组的血清VEGF水平显著升高。在体外实验中，来自纤维化荷瘤动物的NK细胞降低了NPA细胞的增殖。这种降低归因于患有NPA肿瘤的纤维化动物中NK细胞活性的增加。
我们的结果表明，尽管NK细胞在伴有肿瘤的晚期纤维化中受到刺激，但它们可能由于T4和VEGF的分泌而失去了抗肿瘤和抗纤维化活性，这可能解释了慢性HCV患者甲状腺肿瘤风险增加的原因。
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