The effects of GFPT1 on non-small cell lung cancer cells by adjusting the PD-L1 pathway

doi:10.11958/20260080

Abstract

Abstract:

Objective To discuss the effects of glutamine fructose-6-phosphate transaminase 1 (GFPT1) on the proliferation, apoptosis and immune escape of non-small cell lung cancer cells by adjusting the programmed cell death ligand 1 (PD-L1) pathway. Methods Cancer and adjacent tissue samples of 80 patients with non-small cell lung cancer who underwent surgical treatment were collected, and the expression of GFPT1 was detected by immunohistochemistry. Human non-small cell lung cancer cell line A549 was cultured and divided into the CK group, the sh-NC group, the sh-GFPT1 group, the sh-GFPT1+oe-NC group and the sh-GFPT1+oe-PD-L1 group. EdU staining was performed to detect cell proliferation. Cell apoptosis was measured by flow cytometry. Protein expression levels were measured by Western blot assay. Primary human CD8?T cells were isolated and purified from peripheral blood of healthy volunteers and co-cultured with A549 cells from each group for 48 h. The co-culture supernatant was collected, and CD8?T cells were purified and recovered via Ficoll density-gradient centrifugation. Trypan blue staining was used to detect the survival rate of the recovered CD8? T cells from co-culture. Enzyme-linked immunosorbent assay was performed to measure the levels of transforming growth factor-β (TGF-β), interleukin-10 (IL-10), interferon-γ (IFN-γ) and PD-L1 in the co-culture medium. Results The positive rate of GFPT1 was higher in cancer tissue than that in adjacent non-cancerous tissue, and the positive rate of GFPT1 was also higher in cancer tissue from stage Ⅲ—Ⅳ patients compared to those of stage Ⅰ—Ⅱ patients (P<0.01). The positive rate of EdU, the expressions of proliferating cell nuclear antigen (PCNA), Ki-67 proliferation antigen (Ki-67), B-cell lymphoma 2 (Bcl-2) and PD-L1 were lower in the sh-GFPT1 group than those in the CK group and the sh-NC group, while the apoptosis rate and the expression of Bcl-2-associated X protein (Bax) were higher (P < 0.05). The above indicators showed the opposite changes in the sh-GFPT1+oe-PD-L1 group compared with the sh-GFPT1 group and the sh-GFPT1+oe-NC group. After co-culture with CD8⁺T cells, the survival rate of CD8?T cells and IFN-γ level were higher in the sh-GFPT1 group than those in the CK group and the sh-NC group, while the levels of TGF-β, IL-10 and PD-L1 were lower (P<0.05). The survival rate of CD8?T cells and IFN-γ level were lower in the sh-GFPT1+oe-PD-L1 group than those in the sh-GFPT1 group and the sh-GFPT1+oe-NC group, while the levels of TGF-β, IL-10 and PD-L1 were higher (P < 0.05). Conclusion GFPT1 is highly expressed in non-small cell lung cancer tissue. Silencing the expression of GFPT1 can inhibit PD-L1 expression, thereby inhibiting the proliferation and immune escape of non-small cell lung cancer cells and promoting their apoptosis.

Key words: carcinoma, non-small-cell lung, glutamine-fructose-6-phosphate transaminase (isomerizing), B7-H1 antigen, cell proliferation, apoptosis, tumor escape

CLC Number:

R734.2

Figures/Tables 9

References 26

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组别	EdU阳性率	细胞凋亡率	CD8⁺T细胞存活率
CK组	26.41±2.67	6.16±1.76	33.49±5.78
sh-NC组	26.39±2.65	6.14±1.74	33.51±5.82
sh-GFPT1组	9.14±2.47^ab	19.67±1.97^ab	60.35±6.05^ab
sh-GFPT1+oe-NC组	9.15±2.50	19.68±1.98	60.36±6.07
sh-GFPT1+oe-PD-L1组	15.78±2.59^cd	12.79±1.85^cd	46.17±5.96^cd
F	67.956^＊＊	79.084^＊＊	30.709^＊＊

组别	EdU阳性率	细胞凋亡率	CD8⁺T细胞存活率
CK组	26.41±2.67	6.16±1.76	33.49±5.78
sh-NC组	26.39±2.65	6.14±1.74	33.51±5.82
sh-GFPT1组	9.14±2.47^ab	19.67±1.97^ab	60.35±6.05^ab
sh-GFPT1+oe-NC组	9.15±2.50	19.68±1.98	60.36±6.07
sh-GFPT1+oe-PD-L1组	15.78±2.59^cd	12.79±1.85^cd	46.17±5.96^cd
F	67.956^＊＊	79.084^＊＊	30.709^＊＊

组别		PCNA		Ki-67
CK组		0.72±0.08		0.76±0.08
sh-NC组		0.71±0.07		0.78±0.09
sh-GFPT1组		0.35±0.04^ab		0.52±0.05^ab
sh-GFPT1+oe-NC组		0.37±0.05		0.53±0.06
sh-GFPT1+oe-PD-L1组		0.59±0.06^cd		0.69±0.07^cd
F		50.716^＊＊		18.153^＊＊
组别	Bcl-2		Bax		PD-L1
CK组	0.69±0.07		0.35±0.05		0.95±0.10
sh-NC组	0.70±0.08		0.33±0.04		0.97±0.11
sh-GFPT1组	0.43±0.05^ab		0.66±0.08^ab		0.71±0.07^ab
sh-GFPT1+oe-NC组	0.41±0.04		0.64±0.07		0.72±0.08
sh-GFPT1+oe-PD-L1组	0.57±0.06^cd		0.50±0.06^cd		0.83±0.09^cd
F	30.000^＊＊		38.100^＊＊		10.901^＊＊

组别		PCNA		Ki-67
CK组		0.72±0.08		0.76±0.08
sh-NC组		0.71±0.07		0.78±0.09
sh-GFPT1组		0.35±0.04^ab		0.52±0.05^ab
sh-GFPT1+oe-NC组		0.37±0.05		0.53±0.06
sh-GFPT1+oe-PD-L1组		0.59±0.06^cd		0.69±0.07^cd
F		50.716^＊＊		18.153^＊＊
组别	Bcl-2		Bax		PD-L1
CK组	0.69±0.07		0.35±0.05		0.95±0.10
sh-NC组	0.70±0.08		0.33±0.04		0.97±0.11
sh-GFPT1组	0.43±0.05^ab		0.66±0.08^ab		0.71±0.07^ab
sh-GFPT1+oe-NC组	0.41±0.04		0.64±0.07		0.72±0.08
sh-GFPT1+oe-PD-L1组	0.57±0.06^cd		0.50±0.06^cd		0.83±0.09^cd
F	30.000^＊＊		38.100^＊＊		10.901^＊＊

组别		TGF-β	IL-10
CK组		412.69±42.31		119.55±12.02
sh-NC组		412.63±42.27	119.52±11.98
sh-GFPT1组		216.35±40.95^ab	72.14±11.29^ab
sh-GFPT1+oe-NC组		216.17±40.88	72.17±11.35
sh-GFPT1+oe-PD-L1组		367.28±41.08^cd	93.04±11.61^cd
F		35.534^＊＊	24.863^＊＊
组别	IFN-γ			PD-L1
CK组	185.61±30.73			83.72±8.51
sh-NC组	185.64±30.76			83.69±8.47
sh-GFPT1组	316.43±31.71^ab			51.23±8.16^ab
sh-GFPT1+oe-NC组	316.50±31.75			51.19±8.13
sh-GFPT1+oe-PD-L1组	220.71±30.98^cd			72.65±8.30^cd
F	27.183^＊＊			23.374^＊＊