Abstract

The paper aimed to investigate the effects of up-regulation and down-regulation of Mir-299-3p on the proliferation, migration, invasion and apoptosis of lung adenocarcinoma cell line H1299, and to preliminarily explore the mechanism of Mir-299-3p regulating the biological behavior of lung adenocarcinoma cells. Mir-299-3p inhibitor and Mir-299-3p mimics were transfected into lung adenocarcinoma cell line H1299, and the transfection efficiency was measured by fluorescence microscopy. The successfully transfected lung adenocarcinoma cells were detected of proliferation, migration, invasion, and apoptosis. Mir-299-3p inhibitor and Mir-299-3p mimics were used to down-regulate and up-regulate Mir-299-3p respectively. Western blot was employed to detect the expressions of STAT3, p-STAT3, caspase3, Bcl-2, Bax and MMP-9 proteins in lung adenocarcinoma cells. Small interfering RNA technology was applied to construct a cell line with low expression of PTPRD. WB was used to measure the transfection efficiency to determine whether the down-regulation of PTPRD can partially or completely reverse the proliferation, migration, invasion, and apoptosis of lung adenocarcinoma cells after the down-regulation of Mir-299-3p. It was also determined whether the down-regulation of PTPRD can partially or completely reverse the effect of Mir-299-3p down-regulation on the expressions of p-STAT3, MMP-9, Cleaved-caspase3, Bcl-2 and Bax proteins in lung adenocarcinoma cells. Down-regulation of Mir-299-3p can inhibit the proliferation and invasion of lung adenocarcinoma cell H1299, promote cell apoptosis, inhibit the expression of p-STAT3 and MMP-9, promote the expressions of apoptosis-inducing proteins such as Bax and Cleaved-caspase3, and also inhibit the expression of apoptosis inhibitor protein Bcl-2. The up-regulation of Mir-299-3p worked in the opposite way. PTPRD is the direct target gene of Mir-299-3p. Mir-299-3p may directly target PTPRD to regulate the expressions of p-STAT3, MMP-9, Cleaved-caspase3, Bcl-2 and Bax proteins to affect the biological behavior of lung adenocarcinoma cells.

Highlights

  • At present, the high incidence and high mortality of lung cancer have always threatened the health and lives of the people

  • Mir-299-3p may directly target PTPRD to regulate the expressions of p-STAT3, MMP-9, Cleaved-caspase3, Bcl-2 and Bax proteins to affect the biological behavior of lung adenocarcinoma cells

  • The cellular proteins were extracted from the Mir-299-3p inhibitor, Mir-299-3p inhibitor + siPTPRD and NC groups respectively, and the expression of each protein was detected by Western blot

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Summary

Introduction

The high incidence and high mortality of lung cancer have always threatened the health and lives of the people. Lung cancer incidence and mortality rank first among all malignant tumors in men, and rank second in women. The pathogenesis of lung cancer is very complicated, and the current research has not been able to clarify it. Studies have shown that miRNAs play a vital role in the occurrence of tumors. A number of studies have confirmed that miRNA can suppress or promote cancer in tumors. A miRNA can play different roles in different tumors, with complex mechanism (Linden et al, 2020; Arbour & Riely, 2019). It is expected to provide a new theoretical basis for early diagnosis, targeted therapy and prognosis evaluation of lung adenocarcinoma

CCK8 cell proliferation experiment
The effect of Mir-299-3p on the proliferation ability of H1299 cells
Western blot detection
The effect of Mir-299-3p on the scratch healing ability of H1299 cells
The effect of Mir-299-3p on the migration and invasion ability of H1299 cells
The effect of Mir-299-3p on cell apoptosis
The effect of Mir-299-3p on the expressions of various proteins
Discussion
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