Hsa-miR-654-5p调控结肠癌细胞恶性生物学行为及临床意义研究

王海全; 李朝辉; 孙生安; 王云帅; 石守森

doi:10.16766/j.cnki.issn.1674-4152.003749

Hsa-miR-654-5p调控结肠癌细胞恶性生物学行为及临床意义研究

doi: 10.16766/j.cnki.issn.1674-4152.003749

洛阳市中心医院胃肠外科，河南洛阳 471000

基金项目:

河南省医学科技攻关计划联合共建项目 LHGJ20220939

河南省医学科技攻关计划联合共建项目 LHGJ20220940

详细信息

通讯作者:
王海全，E-mail：1281733563@qq.com

中图分类号: R735.35 R730.43
计量
- 文章访问数: 20
- HTML全文浏览量: 7
- PDF下载量: 1
- 被引次数: 0
出版历程
- 收稿日期: 2023-11-24
- 网络出版日期: 2024-12-31

Research on the regulation of malignant biological behavior of colon cancer cells by Hsa miR-654-5p and its clinical significance

Gastroenterology Department of Luoyang Central Hospital, Luoyang, Henan 471000, China

摘要

摘要: 目的检测结肠癌中Hsa-miR-654-5p表达，分析其对结肠癌细胞恶性生物学行为的影响及临床意义。方法采用qRT-PCR法检测结肠癌细胞(HT-29、Caco-2、SW480、HCT116、LOVO)与人正常结肠上皮细胞(HCoEpiC)，2020年1月—2022年12月在洛阳市中心医院中心实验室收集的90例结肠癌与癌旁组织中Hsa-miR-654-5p表达，分析其与患者临床病理特征及预后的关系。按照简单随机分组法进行实验分组：NC组、miR-con组、Hsa-miR-654-5p过表达组及敲减组(每组n=10)。采用噻唑蓝比色法、克隆形成实验、细胞划痕试验和Transwell实验测定各组细胞增殖、克隆、迁移、侵袭变化，蛋白印迹法检测E-钙黏蛋白、人波形蛋白及神经性钙黏附蛋白表达。结果 Hsa-miR-654-5p在结肠癌细胞及组织中表达量明显增高，且不同Hsa-miR-654-5p表达水平患者肿瘤大小、分化程度、TNM分期及淋巴结转移比较差异均有统计学意义(P＜0.05)，而年龄、性别、病理学类型及癌胚抗原比较差异均无统计学意义(P＞0.05)。与NC组和miR-con组相比，Hsa-miR-654-5p过表达组SW480细胞增殖及克隆能力、细胞迁移率、穿膜细胞数、E-钙黏蛋白、人波形蛋白及神经性钙黏附蛋白表达均显著增高，而敲减组细胞上述指标明显降低，组间差异均有统计学意义(P＜0.05)。Hsa-miR-654-5p高表达患者中位生存期显著低于低表达者(20.44个月vs. 38.16个月，χ²=10.079，P＜0.001)。结论 Hsa-miR-654-5p在结肠癌中高表达，且与患者临床病理特征及预后相关，敲减表达后可抑制结肠癌细胞恶性生物学行为，机制可能与抑制上皮-间充质转化有关。
- 结肠癌 /
- Hsa-miR-654-5p /
- 细胞增殖 /
- 侵袭 /
- 预后
Abstract: Objective Detect the expression of Hsa-miR-654-5p in colon cancer, analyze its impact on the malignant biological behavior of colon cancer cells and its clinical significance. Methods Detect the expression of Hsa-miR-654-5p in colon cancer cells (HT-29, Caco-2, SW480, HCT116, LOVO) and human normal colon epithelial cell (HCoEpiC), as well as in 90 cases of colon cancer and adjacent tissues collected from the central laboratory of Luoyang Central Hospital from January 2020 to December 2022 by qRT-PCR, analyze the relationship between its expression and clinical pathological characteristics and prognosis of patients. Conduct experimental grouping according to the simple random grouping method: NC group, miR-con group, Hsa-miR-654-5p overexpression group and knockdown group (n=10 per group). Cell proliferation, cloning, migration, and invasion changes in each group were measured by thiazole blue colorimetry, clone formation assay, cell scratch assay, and Transwell assay, detected the expression of Epithelia cadherin, human vimentin, and Nervous cadheri by Western blotting. Results The expression level of Hsa-miR-654-5p was significantly increased in colon cancer cells and tissues. There were significant differences in tumor size, differentiation, TNM staging, and lymph node metastasis (P < 0.05), while there were no statistically significant differences in age, gender, pathological type, and carcinoembryonic antigen (P>0.05). The proliferation and clone ability, cell migration rate, number of transmembrane cells, and protein expression of Epithelia cadherin, human vimentin, and Nervous cadherin of SW480 cells in the Hsa-miR-654-5p overexpression group were significantly increased when compared with the NC group and miR-con group, while the knockdown group showed a significant decrease in the above indicators, with significant differences between groups (P < 0.05).The median survival of colon cancer patients with high expression of Hsa-miR-654-5p was significantly lower than those with low expression (20.44 months vs. 38.16 months, χ²=10.079, P < 0.001). Conclusion Hsa-miR-654-5p is highly expressed in colon cancer and significantly correlated with clinicopathological features and prognosis of patients. Knockdown of it can inhibit the malignant biological behavior of colon cancer cells, which may be related to the inhibition of epithelial-mesenchymal transition.
- Colon cancer /
- Hsa-miR-654-5p /
- Cell proliferation /
- Invasion /
- Prognosis

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图 1 Hsa-miR-654-5p在结肠癌中的表达

注：与HCoEpiC组比较，^aP＜0.05；与癌旁组织比较，^bP＜0.05。

Figure 1. Expression levels of Hsa-miR-654-5p in colon cancer

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图 2 Hsa-miR-654-5p表达与结肠癌患者预后间的相关性

Figure 2. Correlation between the expression of Hsa-miR-654-5p and prognosis in colon cancer patients

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图 3 转染Hsa-miR-654-5p对结肠癌细胞增殖及克隆的影响

注：与NC组及miR-con组比较，^aP＜0.05。

Figure 3. Effect of Hsa-miR-654-5p transfection on the proliferation and colony formation of colon cancer cells

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图 4 转染Hsa-miR-654-5p对结肠癌细胞相关蛋白表达的影响

注：与NC组及miR-con组分别比较，^aP＜0.05。

Figure 4. Effect of Hsa-miR-654-5p transfection on the expression of related proteins in colon cancer cells

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表 1 Hsa-miR-654-5p表达与结肠癌患者临床病例资料间的关系(例)

Table 1. Relationship between Hsa--miR-654-5p expression and clinical data in colon cancer patients (cases)

项目	例数	Hsa-miR-654-5p		统计量	P值
项目	例数	低表达(n=14)	高表达(n=76)	统计量	P值
年龄				＜0.001^a	0.999
≥40岁	58	9	49
＜40岁	32	5	27
性别				＜0.001^a	0.999
男性	63	10	53
女性	27	4	23
肿瘤直径				-2.675^b	0.007
＜2 cm	27	9	18
2~5 cm	52	4	48
＞5 cm	11	1	10
病理学类型				0.491^a	0.484
腺癌	79	11	68
其他类型	11	3	8
肿瘤部位				0.414^a	0.999
右半结肠	18	3	15
左半结肠	16	2	14
乙状结肠	46	8	38
降结肠、横结肠	10	1	9
血清CEA				0.468^a	0.494
≤10 ng/mL	33	4	29
＞10 ng/mL	57	10	47
TNM分期				-2.777^b	0.005
Ⅰ	25	8	17
Ⅱ	38	5	33
Ⅲ	27	1	26
分化程度				-3.012^b	0.003
高分化	32	10	22
中分化	29	3	26
低分化	29	1	28
淋巴结转移				7.902^a	0.005
有	37	1	36
无	53	13	40
注：^a为χ²值，^b为Z值。

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表 2 不同组别SW480细胞迁移及侵袭能力变化(x±s)

Table 2. Changes in migration and invasion abilities of SW480 cells across different groups (x±s)

组别	n	细胞迁移率(%)	穿膜细胞数(个)
Hsa-miR-654-5p过表达组	20	14.83±1.99^ab	88.89±7.79^ab
Hsa-miR-654-5p敲减组	20	1.16±0.17^ab	5.26±1.18^ab
NC组	20	4.16±1.51	14.66±2.74
miR-con组	20	3.73±1.09	15.28±2.36
F值		391.943	1 605.640
P值		＜0.001	＜0.001
注：与NC组比较，^aP＜0.05；与miR-con组比较，^bP＜0.05。

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