Role of Cytokines Released During Pyroptosis in Non-Small Cell Lung Cancer

Figures & data

Table 1 Summary of Clinical Characteristics of 110 Lung Cancer Patients

Characteristics		Patient No. (%)
Tumor types	Squamous cell carcinomas	72 (65.5)
	Adenocarcinoma	38 (34.5)
Age	Range	40–81
	Median	65
	>65y	53 (48.2)
	≤65y	57 (51.8)
Gender	Male	85 (77.3)
	Female	25 (22.7)
Pathologic grade	I	27 (24.5)
	II	57 (51.8)
	III	26 (23.6)
TNM stage	Stage I	35 (31.8)
	Stage II	33 (30.0)
	Stage III	40 (36.4)
	Stage IV	2 (1.8)
Lymph nodes	Positive	44 (40.0)
	Negative	66 (60.0)
Vital states	Alive	43 (39.1)
	Dead	67 (60.9)
Tumor size (Size of primary lesion)	≤5 cm	82 (74.5)
	>5 cm	28 (25.5)

Table 2 Comparison of Protein Expressions of GSDME, HMGB1 and Caspase3 and CD8+T Lymphocyte Abundance in Lung Cancer and Paracancerous Tissues

		Tumor Tissue (n(%))	Peritumor Tissue (n(%))	p value
GSDME	Low	58 (44.6)	17 (13.1)	0.002*
	High	52 (40)	3 (2.3)
caspase3	Low	17 (13.1)	13 (10.0)	<0.001*
	High	93 (71.5)	7 (5.4)
HMGB1	Low	68 (52.3)	18 (13.9)	0.002*
	High	42 (32.3)	2 (1.5)
CD8^+T lymphocytes	Low	51 (39.2)	17 (13.1)	<0.001*
	High	59 (45.4)	3 (2.3)

Note: *The p value is statistically significant.

Table 3 The Relationship Between the Protein Expression of GSDME, HMGB1 and Caspase3 and the Abundance of CD8+ T Lymphocytes and the Clinical Parameters of 110 Lung Cancer Patients

Variable	GSDME			Caspase-3			HMGB1			CD8^+T Lymphocytes
	Low	High	P value	Low	High	P value	Low	High	P value	Low	High	P value
Age
>65	27	26	0.718	8	45	0.92	29	24	0.139	21	32	0.172
≤65	31	26		9	48		39	18		30	27
Gender
Male	47	38	0.32	12	73	0.474	51	34	0.469	39	46	0.852
Female	11	14		5	20		17	8		12	13
Pathologic grade
I	13	14	0.752	3	24	0.178	18	9	0.794	12	15	0.824
II	32	25		7	50		35	22		28	29
III	1320	13		7	19		15	11		11	15
TNM stage24
I	13	22	0.166	5	30	0.897	19	16	0.67	14	21	0.837
II	20	13		6	27		21	12		16	17
III	24	16		6	34		27	13		20	20
IV	1	1		0	2		1	1		1	1
Lymphatic invasion
Yes(N1,N2,N3)	31	13	0.002*	7	37	0.914	33	11	0.02*	26	18	0.029*
No(N0)	27	39		10	56		35	31		25	41
Vital states
Alive	15	28	0.003*	4	39	0.153	23	20	0.15	17	26	0.25
Dead	43	24		13	54		45	22		34	33
Tumor size
≤5(T1,T2)	44	38	0.738	13	69	0.843	50	32	0.756	40	42	0.384
>5(T3,T4)	14	14		4	24		18	10		11	17
Expression of GSDME
Low	-	-	-	14	44	0.008*	50	8	<0.001*	40	18	0.000*
High	-	-		3	49		18	34		11	41
Expression of caspase-3
Low	14	3	0.008*	-	-	-	14	3	0.058	11	6	0.099
High	44	49		-	-		54	39		40	53
Expression of HMGB1
Low	50	18	<0.001*	14	54	0.058	-	-	-	44	24	<0.001*
High	8	34		3	39		-	-		7	35
Abundance of CD8^+T lymphocytes
Low	40	11	0.001*	11	40	0.099	44	7	<0.001*	-	-	-
High	18	41		6	53		24	35		-	-

Notes: *The p value is less than 0.05, and the difference in protein expression and CD8+ T lymphocyte abundance between clinicopathological parameters is statistically significant.

Figure 1 (A–C) represent the high expression of GSDME, caspase3 and HMGB1 proteins in lung cancer tissues, respectively. (D) The expression of high abundance of CD8+T lymphocytes in lung cancer tissues. (E–G) are GSDME, caspase3, HMGB1 protein in lung cancer tissue low expression. (H) Low abundance of CD8+T lymphocytes in lung cancer tissues. (I–L) is the distribution of GSDME, caspase3, HMGB1, and CD8+T lymphocytes in paracancerous tissues. In the figure, the black arrows is a high expression image and the red arrows is a low expression image. (Images were acquired at 200 × magnification.).

Table 4 Univariate and Multivariate Analysis of Overall Survival

	Univariate Analysis	Multivariate Analysis
	P	P	HR	95% CI
GSDME	0.004*	0.017*	0.531	0.316–0.893
caspase-3	0.259
HMGB1	0.088
CD8^+T	0.348
Age	0.057
Gender	0.999
TNM stage	0.001*	0.026*	1.65	1.061–2.566
Pathologic grade	0.178
Lymphatic invasion	0.042*	0.468	0.788	0.414–1.501
Tumor size	0.019*	0.456	1.25	0.695–2.249

Note: *The p value is statistically significant.

Abbreviations: CI, confidence interval; HR, hazard ratio.

Figure 2 Kaplan-Meier survival curve. In Kaplan-Meier survival analysis, the expression level of GSDME (A), TNM stage (B), lymph node invasion (C) and tumor size (D) were the factors affecting the prognosis of patients (P=0.003, P=0.000, P=0.016, P=0.038, respectively). The expression levels of HMGB1 (E), caspase3 (G) and the abundance of CD8+T lymphocytes (F) were not used as prognostic indicators for the survival of lung cancer patients (P=0.082, P=0.252, P=0.342, respectively). Multivariate analysis of survival curve showed P =0.001 (H). In the TNM staging image, lines of different colors represent different TNM staging, as shown in the notes next to the image. Kaplan-Meier survival curve of other variables, red curve represents high expression and blue curve represents low expression.

Table 5 Correlation Between GSDME, Caspase3, HMGB1 and CD8+ T Lymphocytes and Other Clinicopathological Parameters of Lung Cancer Patients

Variables	GSDME		Caspase3		HMGB1		Abundance of CD8 ^+ T Lymphocytes
	ρ(rho)	P-value	ρ(rho)	P-value	ρ(rho)	P-value	ρ(rho)	P-value
Age	−0.034	0.721	−0.01	0.921	−0.141	0.142	−0.13	0.175
Gender	0.095	0.325	−0.068	0.479	−0.069	0.474	−0.018	0.854
TNM stage	−0.175	0.067	0.009	0.927	−0.099	0.303	−0.081	0.401
Pathologic grade	−0.014	0.883	−0.15	0.117	0.064	0.505	0.014	0.886
Lymphatic invasion	−0.29	0.002*	−0.01	0.915	−0.222	0.020*	−0.208	0.029*
Vital states	0.286	0.002*	0.136	0.156	0.137	0.152	0.11	0.254
Tumor size	−0.175	0.067	0.019	0.845	−0.03	0.758	0.083	0.389
Expression of GSDME	–	–	0.254	0.007*	0.53	0.000*	0.479	0.000*
Expression of caspase-3	0.254	0.007*	–	–	0.181	0.059	0.157	0.101
Expression of HMGB1	0.53	<0.001*	0.181	0.059	–	–	0.468	<0.001*
Abundance of CD8 ^+ T lymphocytes	0.479	<0.001*	0.157	0.101	0.468	<0.001*	–	–

Note: *The p value is statistically significant.

Abbreviation: ρ(rho), Spearman correlation coefficient.