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Western blot analysis technique in detecting protein expression level

2021-09-03
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Lung cancer is one of the most harmful malignant tumors. To explore the pathogenesis of lung cancer and find new targets for anti-tumor therapy are the hotspots and difficulties in the current research on lung cancer treatment. Studies have found that aquaporin 4 (AQP4) is highly expressed in lung cancer cells, and knocking down AQP4 can significantly reduce the migration ability of lung cancer cells. As a technique widely used in protein detection and analysis, Western blot analysis can detect protein expression levels in cells or extracts. Studies have found that aquaporin 4 can be used as a target for targeted therapy of lung cancer. Targeted therapy is a kind of gene product specifically expressed by tumor cells as a therapeutic target, so as to kill tumor cells to the greatest extent, and try not to have a killing effect on normal cells.

In 1994, researchers cloned a selective water transporter-AQP4 protein from the lung tissue of rats, which belongs to the mercury-insensitive AQP family. The researchers used electron microscopy and immunocytochemistry to detect that AQP4 is located on the top of the stromal side of the trachea, bronchial epithelium, and fibroblasts of the lung tissue. In addition, AQP4 is also present on the stromal side of the columnar epithelium of the trachea and bronchus, and AQP4 is located in the alveolar water. It plays an important role in the transportation and adjustment of airway surface fluid. Exploring the effect of aquaporin 4 on lung cancer cells can provide rich research information for the research of lung cancer targeted therapy.

The Biology Department of Shanghai Medicilon has extensive experience in the field of in vitro biology, through enzyme level determination, cell level determination, cell biology, biochemistry, in vitro isotope determination, stable cell line establishment, gene knockout, RNAi and MicroRNA Technology, etc., provide a complete set of biological services.

Immunohistochemistry, reverse transcription polymerase chain reaction (RT-PCR), or Western blot analysis methods can be used to detect the expression and distribution of AQP4 in tissue samples. Western blot analysis is also called Western blotting. It is a new immunobiochemical technology developed on the basis of gel electrophoresis and solid-phase immunoassay technology. It has both the high resolution of gel electrophoresis and solid-phase immunoassay. High specificity and high sensitivity of the assay. As an effective method for identifying antigens with the help of specific antibodies, it has the advantages of simple method, long-term preservation of specimens, and easy comparison of results. It is widely used in molecular biology and other fields. Medicilon provides new drug development outsourcing services. Its biological department has extensive experience in the field of in vitro biology, through enzyme level determination, cell level determination, cell biology, biochemistry, in vitro isotope determination, stable cell line establishment, and gene knockout In addition, RNAi and MicroRNA technologies, etc., provide a complete set of biological services.

In order to study the effect of aquaporin AQP4 on the proliferation of lung cancer cells and the molecular mechanism, the researchers used the CCK-8 kit to detect the proliferation ability of the tumor cells after transfection of AQP4 siRNAs in the lung cancer cell A549, and the Western Blotting method to detect CCND1 The protein level of CCND1 was detected by Real-time PCR method [1]. Studies have shown that in lung cancer cell A549, knocking down the expression of AQP4 can significantly inhibit the proliferation of tumor cells, while down-regulating the mRNA and protein levels of cyclin CCND1. Therefore, AQP4 may become a potential target for lung cancer treatment.

Some researchers have also explored the influence of aquaporin AQP4 on the migration ability of lung cancer cells [2]. The method is to select lung cancer A549 cell line, use AQP4 small interfering RNA (sjRNAs) for transfection, use Transwell method to detect tumor cell migration ability, and use Western blot method to detect epithelial cadherin (E-cadherin) and neural cadherin (N-cadherin) and vimentin (Vimentin) expression levels, and analyze their effects on epithelial interstitial transformation (EMT). The results showed that knocking down AQP4 can inhibit lung cancer cell A549 migration ability and EMT, up-regulate the expression level of E-cadherin and down-regulate the expression level of N-cadherin and Vimentin (p<0.05). Therefore, inhibiting the expression of AQP4 can effectively reduce the migration ability of lung cancer cells A549 and inhibit EMT. It is speculated that promoting EMT may be the main molecular mechanism of AQP4.

There are many studies on AQP4 protein, so I won’t list them all here. In short, in-depth study of the carcinogenic mechanism of lung cancer and discovery of anti-tumor treatment targets are of great significance for the diagnosis and treatment of lung cancer.

[1] The effect of aquaporin AQP4 on the proliferation of lung cancer cells [J].

[2] The effect of aquaporin 4 on the migration ability of lung cancer cell A549 and its mechanism [J].

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