Role of NID1 in angiogenesis of clear cell renal cell carcinoma

doi:10.11958/20240301

Abstract

Abstract:

Objective To explore the expression of nestin-1 (NID1) in clear cell renal cell carcinoma (ccRCC) and its effect on angiogenesis. Methods The expression of NID1 in ccRCC was analyzed using data from the TCGA database, cell lines and clinical samples. Additionally, the relationship between NID1 and the angiogenesis marker CD31 was investigated. A stable knockdown 786-O cell line with reduced NID1 expression was generated through lentiviral transfection. The conditioned medium obtained from these cells was used to treat human umbilical vein endothelial cells (HUVECs). HUVEC cells were divided into four groups: the DMEM group, the blank group, the NID1 empty vector group and the NID1 knockdown group. Cell proliferation ability was assessed using CCK-8 method, while tube formation ability and migration ability were evaluated through tube formation test and scratch test respectively. A mouse model of dextran hydrogel unloaded/NID1 knockdown tumor cell complex culture system was established and divided into the unloaded NID1 group and the NID1 knockdown group. Immunohistochemical staining was performed to detect the expression of CD31. Results Bioinformatics analysis revealed an increased expression of NID1 in ccRCC tissue. Furthermore, 500 differentially expressed genes positively correlated with elevated levels of NID1 were primarily enriched in angiogenesis-related processes. The results showed a positive correlation between the expression of NID1 and CD31 in cancer tissue. After treatment with conditioned medium derived from the knockdown group, HUVEC cells exhibited weakened migration and tube formation abilities in the empty vector control group (P<0.05), while their proliferation ability remained unchanges compared to the empty vector control group (P>0.05). In vivo experiments using a nude mouse model, the expression of CD31 demonstrated significantly lower levels in the NID1 knockdown group (P<0.05). Conclusion The expression of NID1 is increased in ccRCC, and it can promote angiogenesis.

Key words: carcinoma, renal cell, extracellular matrix, cell proliferation, angiogenesis

CLC Number:

R692.9

Figures/Tables 7

Fig.1 Expression of NID1 in ccRCC cells

Fig.2 Relationship between NID1 and angiogenesis

Fig.3 NID1 expression levels detected in 786-O cells and culture supernatant of each group

Tab.1 Comparison of proliferation ability, tube number and migration healing rate between four groups of cells

组别	细胞增殖能力/ （OD₄₅₀）	成管数/ （个/视野）	迁移愈合率/%
DMEM组	0.83±0.15	1.67±0.58	7.86±0.49
空白组	0.72±0.03	24.67±6.11^a	18.11±3.35^a
NID1空载组	0.84±0.13	22.67±4.04^a	17.05±0.34^a
NID1敲低组	0.90±0.15	5.67±0.58^b	10.87±0.61^b
F	1.107	30.180^＊＊	24.320^＊＊

Fig.4 Effect of NID1 expression on HUVEC tube formation (×10)

Fig.5 Effect of NID1 expression on HUVEC migration

Fig.6 Effect of NID1 expression on angiogenesis in vivo (CD31 staining, ×40)

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