Silencing LMNB1 Contributes to the Suppression of Lung Adenocarcinoma Development

Figures & data

Table 1 Expression Patterns of LMNB1 in LUAD Tissues and Normal Tissues Revealed in Immunohistochemistry Analysis

LMNB1 Expression	Tumor Tissue		Normal Tissue
	Cases	Percentage	Cases	Percentage
Low	44	51.2%	72	96%
High	42	48.8%	3	4%

Note: P<0.001.

Table 2 Relationship Between LMNB1 Expression and Tumor Characteristics in Patients with LUAD

Features	No. of Patients	LMNB1 Expression		P-value
		Low	High
All patients	86	44	42
Age (years)				0.391
≤61	43	20	23
>61	43	24	19
Gender				0.269
Male	48	22	26
Female	38	22	16
Tumor size				0.119
<4 cm	36	22	14
≥4 cm	50	22	28
Lymph node positive				0.006
≤1	47	30	17
>1	36	12	24
Grade				0.356
I	3	2	1
II	56	30	26
III	27	12	15
Stage				0.012
1	25	18	7
2	16	8	8
3	39	14	25
4	1	1	0
T infiltrate				0.187
T1	16	11	5
T2	49	23	26
T3	15	9	6
T4	6	1	5
Lymphatic metastasis (N)				0.002
N0	36	24	12
N1	14	7	7
N2	16	5	11
N3	4	0	4
Expression of EGFR (FISH)				0.617
Negative	69	37	32
Positive	11	5	6

Table 3 Relationship Between LMNB1 Expression and Tumor Characteristics in Patients with LUAD Analyzed by Spearman Rank Correlation Analysis

Tumor Characteristics	Index
Stage	Pearson's correlation	0.279
	Significance (two tailed)	0.012
	n	81
Lymphatic metastasis (N)	Pearson's correlation	0.365
	Significance (two tailed)	0.002
	n	70
Lymph node positive	Pearson's correlation	0.302
	Significance (two tailed)	0.005
	n	83

Figure 1 Identification of LMNB1 as potential tumor promotor in LUAD. (A) RNA-seq was performed to identify differentially expressed genes (DEGs) in LUAD tumor tissues and normal tissues. (B) Endogenous expression of several selected DEGs was detected by qPCR in NCI-H1299 cells. (C) Celigo cell counting assay was utilized to evaluate the inhibition of cell proliferation by knockdown of the selected DEGs. (D, E) The expression of LMNB1 in LUAD tumor tissues and normal tissues was detected by qPCR (D) and immunohistochemistry analysis (E). (F) Kaplan–Meier survival analysis was performed to show the correlation between LMNB1 expression and prognosis of LUAD patients. *P<0.001.

Figure 2 Depletion of LMNB1 inhibits cell proliferation and migration, induces cell apoptosis and cell cycle arrest. (A) qPCR and Western blotting were performed to assess the knockdown efficiency of LMNB1 in LUAD cells. (B) Effects of LMNB1 knockdown on cell proliferation was determined by Celigo cell counting assay. (C, D) Influence of LMNB1 depletion on cell apoptosis and cell cycle distribution was detected by flow cytometry. (E, F) Impacts of LMNB1 knockdown on LUAD cell migration was examined by wound-healing (E) and transwell (F) assays. *P<0.05, **P<0.01, ***P<0.001.

Figure 3 Exploration of mechanism by which LMNB1 regulates LUAD. (A, B) Human apoptosis antibody array was performed to identify differentially expressed apoptosis-related proteins in NCI-H1299 cells with or without LMNB1 knockdown. (C) Western blotting was performed to detect the changes in expression of proteins in cancer-related signaling pathways. *P<0.05, **P<0.01.

Figure 4 LMNB1 knockdown reduces tumorigenesis of LUAD cells in vivo. (A) Tumor volume was calculated based on tumor size measured at indicated time intervals. Inset: photo of the removed xenografts. (B) In vivo imaging was performed to evaluate tumor growth. (C) The total intensity of bioluminescence was scanned to represent the tumor burden. (D) Tumor weight was measured after sacrificing mice. (E) The expression of Ki67 in tumor sections was detected by immunohistochemistry analysis. *P<0.01, **P<0.001.