Journal of Shanghai Jiao Tong University (Medical Science) ›› 2022, Vol. 42 ›› Issue (5): 578-582.doi: 10.3969/j.issn.1674-8115.2022.05.004

• Basic research • Previous Articles     Next Articles

Effect of altered expression of long non-coding RNA-B230352I09 on proliferation and cycle of H9C2 cardiomyocytes

XU Feixiang(), WANG Sheng, XUE Mingming, TONG Chaoyang, CHEN Yumei()   

  1. Department of Emergency Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China
  • Received:2022-02-14 Accepted:2022-05-16 Online:2022-05-28 Published:2022-05-28
  • Contact: CHEN Yumei;
  • Supported by:
    National Natural Science Foundation of China(81800230)

Abstract: Objective

·To investigate the effect of altered expression of long non-coding RNA (lncRNA)-B230352I09 on proliferation and cycle of H9C2 cardiomyocytes.


·The lncRNA-B230352I09 overexpression vector (pcDNA-B230352I09) and the negative control vector pcDNA-negative control group (NC) were constructed and transfected into H9C2 cardiomyocytes with the Lipofectamine 3000 (Lipo3000) transfection solution. The expression of lncRNA-B230352I09 was measured by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) to verify its transfection efficiency. The H9C2 cardiomyocytes were divided into blank control group, pcDNA-NC group, and lncRNA-B230352I09 overexpression group. Cell counting kit-8 (CCK8) was used to measure the absorbance (optical density, OD) of H9C2 cardiomyocytes at a wavelength of 450 nm and draw a growth curve; 5-ethynyl-2'-deoxyuridine (EdU) was used to label proliferating H9C2 cardiomyocytes, the numbers of which were observed by fluorescence microscopy. Cardiomyocyte cycle was assessed by flow cytometry and cycle-related genes expression was measured by RT-PCR.


·Compared with the pcDNA-NC group, the expression level of lncRNA-B230352I09 was significantly higher in the lncRNA-B230352I09 overexpression group (P=0.000), suggesting that the lncRNA-B230352I09-transfected H9C2 cardiomyocyte model was successfully constructed. Compared with the pcDNA-NC group, the CCK8 assay showed that lncRNA-B230352I09 overexpression significantly increased the OD of H9C2 cardiomyocytes and promoted the proliferation of H9C2 cardiomyocytes with a significant time-dependent enhancement of cell proliferation (24 h: P=0.000; 48 h: P=0.000; 72 h: P=0.001). Fluorescence microscopy revealed that the proportion of EdU-labeled positive H9C2 cardiomyocytes was significantly increased in the lncRNA-B230352I09 overexpression group compared to the pcDNA-NC group. Flow cytometric analysis showed a significant increase in the proportion of H9C2 cardiomyocytes in S phase and a decrease in G1 phase in the lncRNA-B230352I09 overexpression group compared to the pcDNA-NC group (P=0.000). RT-PCR showed that the mRNA expression levels of cyclin D1 and cyclin dependent protein kinase 1 (CDK1) in the lncRNA-B230352I09 overexpression group were significantly higher compared with the pcDNA-NC group (P=0.000).


·lncRNA-B230352I09 can enhance myocardial proliferative capacity by regulating cyclin D1 and CDK1 to promote the entry of cardiomyocytes to into S phase.

Key words: H9C2 cardiomyocyte, lncRNA-B230352I09, overexpression, cell proliferation, cell cycle

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