The study of a single BGC823 cell using Fourier transform infrared microspectroscopic imaging

doi:10.1016/j.saa.2011.05.031

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy

Volume 79, Issue 5, September 2011, Pages 1660-1662

https://doi.org/10.1016/j.saa.2011.05.031 Get rights and content

Abstract

In order to investigate gastric cancer at cellular and sub-cellular level, a single human gastric adenocarcinoma BGC823 cell was studied by an infrared microscope equipped with a focal plane array (FPA) detector. The spectra showed difference between the nucleus and the endoplasmic reticulum (ER) of the BGC823 cell. The peak of v_asPO₂⁻ was shifted to a higher wavenumber at the nucleus compared with that at the ER. The height ratios of 2954 cm⁻¹/2922 cm⁻¹ (CH₃/CH₂) and 1088 cm⁻¹/1539 cm⁻¹ (DNA/amide II) of the nucleus were significantly higher than those of the ER. Furthermore, chemical images reveal the intensity distributions of lipids, proteins and DNA of the single BGC823 cell, and the intense absorptions of proteins and DNA were observed in the nuclear region of the cell while the intense absorption of lipids was found in the ER region of the cell. The Fourier transform infrared (FTIR) microspectroscopic imaging result indicates the study of the single gastric cancer cell at sub-cellular level can be beneficial for knowing gastric cancer more which will be of great importance for the study and diagnosis of gastric cancer. The result also suggests that FPA is a useful tool in the study of a single cell and may be a powerful tool for study and diagnosis of gastric cancer.

Graphical abstract

Highlights

► A single BGC823 cell was studied by IR microspectroscopic imaging with FPA detector. ► The spectra showed difference between the nucleus and the ER of the BGC823 cell. ► Chemical images revealed lipids, proteins and DNA distributions of the BGC823 cell. ► FPA may be useful tool for study and diagnosis of gastric cancer.

Introduction

Since infrared microspectroscopy and imaging is a powerful tool for detecting biological changes in diseased cells and tissues during the process of carcinogenesis, it is widely used for cancer research. For instance, lung cancer [1], cervical cancer [2], gastric cancer [3], [4] were examined by infrared microspectroscopy. The study of a single cancer cell by infrared microspectroscopy can provide particular information of cancer cells at the cellular and sub-cellular level, but it is difficult to study such a small single cancer cell using infrared microspectroscopy with conventional global source.

Although gastric cancer is a common malignancy in the world, there are few detailed studies on gastric cancer cells by infrared spectroscopy. Recently, Fujioka et al. compared infrared spectra of normal gastric mucosal epithelial cells and three gastric cancer cells (AGS, SNU-1 and NCI-N87) [5], and Du et al. used infrared spectroscopy to study gastric cancer cell line SGC7901 [6]. The above studies were based on infrared spectroscopy but not on infrared microspectroscopy, and the results were the average of a great deal cells but not the single cell. To our knowledge, there is no report on studying the single gastric cancer cell with infrared microspectroscopy and imaging till now although it is of great importance to achieve sub-cellular spatial resolution.

Fortunately, focal plane array (FPA) detector, including series of small infrared detector elements, can obtain high spatial resolution [7]. Now FPA is used for study of human hair tissue [7], benign breast tumor tissue [8], human ovarian carcinoma cells SK-OV-3 [9], etc. In this paper, BGC823, a poorly differentiated gastric cancer cell line, was investigated by FPA. The goal is to investigate the possibility of studying the single gastric cancer cell by FPA and provide some useful information for further understanding of gastric cancer's pathogenesis.

Section snippets

Cell culture

Human gastric adenocarcinoma cell line BGC823 (poorly differentiated) was purchased from The Cell Bank of Chinese Academy of Sciences. Cells were cultured upon sterilized BaF₂ and incubated in DMEM medium (Gibco BRL, USA) supplemented with 10% heat-inactivated fetal bovine serum (Sijiqing Biotechnology Co., China), penicillin (100 unit/mL) and streptomycin (100 μg/mL) at 37 °C in a 5% CO₂ humidified atmosphere. After 2 h, BaF₂ window was washed with distilled water and the cells were fixed with 4%

Spectrum of the single BGC823 cell

There were several main bands in the average spectrum of the single BGC823 cell (Fig. 1): 3283 cm⁻¹ (N–H stretching of amide A and O–H stretching of carbohydrates) [10] and 3059 cm⁻¹ (N–H stretching of amide B) [11], 2954 cm⁻¹ and 2869 cm⁻¹ (antisymmetric and symmetric stretching of CH₃) [12], 2922 cm⁻¹ and 2852 cm⁻¹ (antisymmetric and symmetric stretching of CH₂) [12], 1739 cm⁻¹ (C double bond O stretching) [8], 1644 cm⁻¹ (the amide I) and 1539 cm⁻¹ (the amide II) [8], 1451 cm⁻¹ (δasCH₃ and δCH₂ vibrations) and 1400

Conclusion

Infrared microspectroscopic imaging with FPA can help us reveal the BGC823 cell at the sub-cellular level. For the nucleus of the BGC823 cell, the v_asPO₂⁻ band (1244 cm⁻¹) was shifted to higher wavenumber compared with that of the ER (1232 cm⁻¹). The band shift indicates the structure change between the nucleus and the ER. The significant differences of H1088/H1539 and H2954/2922 between the nucleus and the ER may suggest the changes of lipid chains and the content increase of DNA at the nucleus.

Acknowledgments

The authors are grateful for the financial support from the 985 Project of the State Ministry of Education, the National Natural Science Foundation of China (10675113, 10734070, 10974191) and the Knowledge Innovation Program of the Chinese Academy of Sciences (KJCX2-YW-N43).

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The study of a single BGC823 cell using Fourier transform infrared microspectroscopic imaging

Abstract

Graphical abstract

Highlights

Introduction

Section snippets

Cell culture

Spectrum of the single BGC823 cell

Conclusion

Acknowledgments

Anal. Biochem.

Cancer Lett.

Vib. Spectrosc.

Cancer Epidemiol.

J. Mol. Struct.

Cancer Detect. Prev.

Vet. Microbiol.

Biophys. Chem.

J. Mol. Struct.