Impact of ultrasound microbubbles-mediated miR-545-3p delivery on CRC proliferation， apoptosis，and invasion via HOXA3

doi:10.3877/cma.j.issn.2095-5782.2025.02.008

Abstract

Abstract:

Objective

To investigate the impacts of miR-545-3p mediated by ultrasound microbubbles on the proliferation， apoptosis， and invasion of colorectal cancer （CRC） cells by regulating HOXA3.

Methods

Real-time fluorescence quantitative polymerase chain reaction （qRT-PCR） was used to detect the mRNA expression levels of miR-545-3p and homeobox gene A3 （HOXA3） in SW620，SW480， LoVo， CACO-2， and the normal colon epithelial cell line NCM460； SW480 cells with good growth were divided into blank group， microbubble group， miR group， overexpression miR-545-3p group，microbubble+miR group， microbubble+overexpression miR-545-3p group， microbubble+overexpression miR-545-3p+empty vector （pcDNA6.2/nLumio-DEST， pcDNA） group， and microbubble+overexpression miR-545-3p+HOXA3 group.qRT-PCR was used to detect the expression of miR-545-3p and HOXA3 in each group， MTT was used to detect cell proliferation， flow cytometry was used to detect cell apoptosis，Transwell assay was used to detect cell invasion， and Western blotting was used to detect HOXA3 and proliferating cell nuclear antigen （PCNA）.Gene expression levels of PCNA， matrix metalloproteinase 9（MMP-9）， and Bcl-2-associated X protein （Bax） were measured.

Results

The expression of miR-545-3p was down-regulated and HOXA3 mRNA expression was up-regulated in CRC cells （P＜0.05）， and the changes were most significant in SW480 cells， which were selected for follow-up research.The luciferase activity of miR-545-3p co-transfected with HOXA3-WT was lower than that of miR NC co-transfected with HOXA3-WT （P＜0.05）.Overexpression of miR-545-3p can upregulate miR-545-3p expression，downregulate HOXA3 expression， inhibit cell proliferation， invasion and expression of PCNA and MMP-9 proteins， and promote cell apoptosis and Bax protein expression （P＜0.05）.Overexpression of miR-545-3p in the microbubble+miR-545-3p group upregulates miR-545-3p expression， downregulates HOXA3 expression， inhibits cell proliferation， invasion， and expression of PCNA and MMP-9 proteins， and promotes cell apoptosis and Bax protein expression more effectively than overexpression of miR-545-3p alone（P＜0.05）.The tumor weight of the miR-545-3p microbubble group in nude mice was significantly lower than that of the control group， microbubble group， and miR microbubble group （P＜0.05）.

Conclusion

miR-545-3p mediated by ultrasound microbubbles can inhibit the proliferation and invasion of CRC cells and induce their apoptosis by down-regulating HOXA3， providing a new drug delivery method and potential target for clinical treatment of CRC， and providing a basis for molecular targeted therapy of CRC.

Key words: Ultrasound microbubbles, Colorectal cancer, miR-545-3p, HOXA3

Siqi Li, Hanwei Chen, Qiong Ye, Jingen Luo. Impact of ultrasound microbubbles-mediated miR-545-3p delivery on CRC proliferation， apoptosis，and invasion via HOXA3[J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2025, 13(02): 143-151.

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URL: https://zhjrfsxdzzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-5782.2025.02.008

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Figures/Tables 14

References 22

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组别	例数	miR-545-3p	HOXA3mRNA
NCM460组	6	0.95±0.10	1.03±0.11
SW620组	6	0.72±0.08^a	1.45±0.15^a
LoVo组	6	0.68±0.07^a	1.57±0.16^a
CACO-2组	6	0.63±0.07^a	1.53±0.16^a
SW480组	6	0.42±0.05^a	2.13±0.22^a

组别	例数	miR-545-3p	HOXA3mRNA
NCM460组	6	0.95±0.10	1.03±0.11
SW620组	6	0.72±0.08^a	1.45±0.15^a
LoVo组	6	0.68±0.07^a	1.57±0.16^a
CACO-2组	6	0.63±0.07^a	1.53±0.16^a
SW480组	6	0.42±0.05^a	2.13±0.22^a

组别	例数	荧光素酶相对活性
miR NC+HOXA3-WT组	6	1.08±0.11
过表达miR-545-3p+HOXA3-WT组	6	0.59±0.06^a
miR NC+HOXA3-MUT组	6	1.03±0.11
过表达miR-545-3p+HOXA3-MUT组	6	1.07±0.12

组别	例数	荧光素酶相对活性
miR NC+HOXA3-WT组	6	1.08±0.11
过表达miR-545-3p+HOXA3-WT组	6	0.59±0.06^a
miR NC+HOXA3-MUT组	6	1.03±0.11
过表达miR-545-3p+HOXA3-MUT组	6	1.07±0.12

组别	例数	miR-545-3p	HOXA3 mRNA	HOXA3/β-actin
空白组	6	1.02±0.11	0.96±0.10	0.87±0.09
微泡组	6	0.98±0.10	1.07±0.12	0.83±0.09
miR组	6	1.08±0.12	1.01±0.11	0.94±0.10
过表达miR-545-3p组	6	1.65±0.18^abc	0.64±0.07^abc	0.52±0.06^abc
微泡+miR组	6	1.06±0.11	1.03±0.11	0.88±0.09
微泡+过表达miR-545-3p组	6	2.18±0.22^de	0.37±0.04^de	0.34±0.04^de
微泡+过表达miR-545-3p+pcDNA组	6	2.14±0.22	0.34±0.03	0.32±0.04
微泡+过表达miR-545-3p+HOXA3组	6	2.09±0.21	0.72±0.08^f	0.66±0.07^f

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