MiR-374 promoting proliferation and invasion of breast cancer cells by targeting and down-regulating TRIM35

doi:10.3969/j.issn.1673-8640.2023.09.002

Abstract

Abstract:

Objective To investigate the role of miR-374 in breast cancer and the relationship between miR-374 and tripartite motif-containing protein 35（TRIM35）. Methods A total of 42 patients with breast cancer were enrolled，and cancer tissues and adjacent tissues（>2 cm away from tumor resection margin） were collected，and the data of patients were collected as well. The expression of miR-374 and TRIM35 were determined in cancer tissues and adjacent tissues. Breast cancer cell line MCF-7 was classified into 3 groups according to different treatment methods，including control group（without any treatment），interference group（adding 100 nmol/L miR-374 inhibitor） and mock-vehicle group（adding 100 nmol/L negative control sequence）. The effects of miR-374 on the proliferative activity and invasiveness of breast cancer cell line MCF-7 were evaluated by cell proliferation assay and Transwell assay. Dual luciferase reporter gene assay was used to determine the interaction between miR-374 and TRIM35. Results The expression of miR-374 in cancer tissues was higher than that in adjacent tissues（P<0.001）. TRIM35 relative expression in cancer tissues was lower than that in adjacent tissues（P<0.001）. The miR-374 relative expression had statistical significance with different breast tumor sizes，distant metastasis and lymphatic metastasis（P<0.05）. There was no statistical significance for miR-374 relative expression with different ages and TNM stages（P>0.05）. The relative expression of miR-374 in interference group was lower than those in control group and mock-vehicle group（P<0.001）. The cell proliferation rate and the number of invasive cells in interference group were lower than those in mock-vehicle group and control group（P<0.001）. TRIM35 relative expressions were higher than those in mock-vehicle group and control group（P<0.001）. Dual luciferase reporter gene assay confirmed that miR-374 directly inhibited the transcriptional expression of target gene TRIM35. Conclusions MiR-374 interacts with TRIM35，and miR-374 promotes the proliferation and invasion of breast cancer cells by targeting and down-regulating the expression of TRIM35.

Key words: MicroR-374, Tripartite motif-containing protein 35, Breast cancer, Cell proliferation, Cell metastasis

CLC Number:

R393

WANG Rong, XING Lianxiang, HUANG Keliang, LI Xin. MiR-374 promoting proliferation and invasion of breast cancer cells by targeting and down-regulating TRIM35[J]. Laboratory Medicine, 2023, 38(9): 812-817.

Figures/Tables 8

References 24

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基因名称	引物序列（5'~3'）	产物长度/bp
TRIM35	F：GAGGTGCTGGCACATGAGAT	112
	R：GCTCCATGGTGCAGAAGAGT
GAPDH	F：CACCCACTCCTCCACCTTTG	110
	R：CCACCACCCTGTTGCTGTAG
miR-374	F：ATATAATACAACCTGCTAAGTG	134
	R：GCAGGGTCCGAGGTATTC
U6	F：CTCGCTTCGGCAGCACA	94
	R：AACGCTTCACGAATTTGCGT

基因名称	引物序列（5'~3'）	产物长度/bp
TRIM35	F：GAGGTGCTGGCACATGAGAT	112
	R：GCTCCATGGTGCAGAAGAGT
GAPDH	F：CACCCACTCCTCCACCTTTG	110
	R：CCACCACCCTGTTGCTGTAG
miR-374	F：ATATAATACAACCTGCTAAGTG	134
	R：GCAGGGTCCGAGGTATTC
U6	F：CTCGCTTCGGCAGCACA	94
	R：AACGCTTCACGAATTTGCGT

临床病理特征	例数	miR-374
年龄
<60岁	16	3.52±1.68
≥60岁	26	3.30±1.64
t值		0.389 1
P值		0.678 4
肿瘤大小
<3 cm	18	2.64±1.20
≥3 cm	24	3.81±1.67
t值		2.148 3
P值		0.023 2
TNM分期
T1~T2	26	3.40±1.51
T3~T4	16	3.29±1.63
t值		0.532 6
P值		0.551 4
淋巴转移
无	14	3.80±1.55
有	28	2.32±1.07
t值		7.724 2
P值		0.002 1
远隔转移
无	31	2.49±1.54
有	11	3.59±1.49
t值		1.934 5
P值		0.034 9

临床病理特征	例数	miR-374
年龄
<60岁	16	3.52±1.68
≥60岁	26	3.30±1.64
t值		0.389 1
P值		0.678 4
肿瘤大小
<3 cm	18	2.64±1.20
≥3 cm	24	3.81±1.67
t值		2.148 3
P值		0.023 2
TNM分期
T1~T2	26	3.40±1.51
T3~T4	16	3.29±1.63
t值		0.532 6
P值		0.551 4
淋巴转移
无	14	3.80±1.55
有	28	2.32±1.07
t值		7.724 2
P值		0.002 1
远隔转移
无	31	2.49±1.54
有	11	3.59±1.49
t值		1.934 5
P值		0.034 9