Ultrasound-targeted microbubble destruction mediated miR-492 inhibitor suppresses the tumorigenesis in non-small cell lung cancer

Figures & data

Figure 1. Expression of miR-492 in NSCLC tissues and cell lines. (A) The expression of miR-492 was increased in NSCLC tissues. (B) The expression level of miR-492 was increased in four NSCLC cell lines compare with normal NHBE cell lines. (C) NSCLC patients with high miR-492 expression had worse survival compared with low miR-492 expression, illustrating that NSCLC patients with high miR-492expression were associated with worse prognostic survival (Log-rank p = .0007) (***p < .001).

Table 1. Clinicopathological characteristics in patients with NSCLC.

Features	Total (n = 96)	miR-492		p-Value
		Low (n = 46)	High (n = 50)
Age (years)				.963
≤60	29	14	15
>60	67	32	35
Gender				.629
Female	40	18	22
Male	56	28	28
Smoking				.936
No	33	16	17
Yes	63	30	33
Tumour size (cm)				.039
≤3	50	29	21
>3	46	17	29
Histological type				.497
Adenocarcinoma	55	28	27
Squamous cell carcinoma	41	18	23
Differentiation				.055
Well and moderate	55	31	24
Poor	41	15	26
TNM				.006
I–II	55	33	22
III–IV	41	13	28
Lymph node metastasis				.002
Negative	55	34	21
Positive	41	12	29

NSCLC: non-small cell lung cancer.

Table 2. Multivariate Cox regression analysis to adjust the association of miR-492 with NSCLC patients’ survival prognosis.

Variables	HR	95% CI	p-Value
Age	1.119	0.712–1.554	.423
Gender	1.208	0.769–1.637	.379
Smoking	1.375	0.894–1.981	.121
Tumour size	1.224	0.988–1.380	.096
Histological types	1.095	0.634–1.448	.557
Differentiation	1.427	0.976–1.909	.078
TNM stage	2.395	1.569–4.337	.013
Lymph node metastasis	1.875	1.347–2.888	.029
miR-492	2.994	1.815–4.894	.006

NSCLC: Non-small cell lung cancer; TNM: Tumour node metastasis.

Figure 2. miR-492 inhibitor suppresses cell proliferation, migration and invasion of NSCLC cells. (A) After transfection with miR-492 inhibitors, miR-492 expression levels in A549 and PC9 cell lines decreased compared with inhibitor NC (all p < .001). (B, C) Silencing of miR-492 suppressed cell proliferation in A549 and PC9 cell lines compared with inhibitor NC, and the difference gradually became significant with time (48 h: all p < .01; 72 h: all p < .001). (D, E) Silencing of miR-492 significantly inhibited cell migration and invasion in A549 and PC9 cell lines compared with inhibitor NC. Transwell images observed by inverted microscopy showed that the number of cell migration and invasion was the least in A549 and pC9 cells transfected with miR-492 inhibitor (all p < .001) (**p < .01; ***p < .001).

Figure 3. UTMD enhances cell transfection efficiency of miR-492. (A, B) UTMD could significantly enhance the inhibition of miR-492 inhibitor on the expression level of miR-4284 in A549 and PC9 cell lines, and the efficiency of transfection increased (***p < .001; ^#p < .05; ^##p < 0.01; *compare with control, ^##compare with miR-492 inhibitor).

Figure 4. Effects of UTMD-mediated miR-492 inhibitor transfection on NSCLC cell proliferation, migration and invasion. (A) UTMD-mediated miR-492 inhibitor further inhibited cell proliferation compared with transfection of miR-492 inhibitor alone. (B, C) UTMD-mediated miR-492 inhibitor further inhibited cell migration and invasion compared with a single miR-492 inhibitor (*p < .05; **p < .01; ***p < .001; ^#p < .05; *compare with control, ^#compare with miR-492 inhibitor).

Figure 5. Target genes of miR-492 predicted by the miRWalk database.

Data availability statement

The data used to support the findings of this study are available from the corresponding author upon reasonable request.