1.Absorption and elimination of photofrin-Ⅱ in human immortalization esophageal epithelial cell line SHEE and its malignant transformation cell line SHEEC
Gao SHE-GAN ; Wang LI-DONG ; Feng XIAO-SHAN ; Qu ZHI-FENG ; Shan TAN-YOU ; Xie XUAN-HU
Chinese Journal of Cancer 2009;28(12):1248-1254
Background and Objective:The mechanism of tumor tissues selectively uptaking the photosensitizer in photodynamic therapy(PDT)is still unclear.This study was to investigate the affinity of tumor cells to the photosensitizer photofrin-Ⅱ. Methods: Ultraviolet spectrophotometer was applied to measure the absorption spectra of various cell culture media.The fluorescence spectrum of photofrin-Ⅱ was determined by spectrofluorometer.The absorption and eIimination condition of photofrin-Ⅱ were detected in immortalized human esophageal epithelial cell line SHEE and its malignant transformation cell line SHEEC.Results:The maximum excitation wavelength of fluorescence for photofrin-Ⅱ was(395.0±0.5) nm, and the maximum emission wavelength of that was (634.1±0.5) nm.The laser at the wavelength of 630 nm used in this experiment could permeate various types of cell culture media.There was no significant difference in the absorption and elimination of photofrin-Ⅱ between SHEE and SHEEC at the same concentration and time.The absorption of photofrin-Ⅱ in SHEE and SHEEC increased with the increase in photofrin-Ⅱ concentration and duration. and reached the platform at the concentration of 30 μg/mL and a time point of 150 min.respectively.The photofrin-Ⅱ contents of SHEE and SHEEC showed a slight change after 15-30 min, and diminished rapidly after 30 min.Conclusion:High photosensitizer concentration in tumor tissues may be not correlated with the affinity of tumor cells.
2.Expression and clinical significance of KiSS-1 and E-cadherin in gastric cardia carcinoma.
Zhan-hui WANG ; Xiao-shan FENG ; Gong-ping WANG ; She-gan GAO ; Jian-qiang MI ; Jiang-Tao SUN ; Zhi-feng QU ; Jing HAN ; Luo ZHANG
Chinese Journal of Gastrointestinal Surgery 2007;10(4):380-382
OBJECTIVETo investigate the expression and clinical significance of KiSS- 1 and E- cadherin in gastric cardia carcinoma and the correlation between the two proteins.
METHODSThe expression of KiSS- 1 and E- cadherin in 80 patients with gastric cardia carcinoma and 20 patients with normal gastric cardia epithelium was detected by immunohistochemical technique.
RESULTSThe expression of KiSS- 1 was negatively correlated with lymphatic metastasis and clinical stage (P < 0.05), but not correlated with the cancer differentiation (P < 0.05). The expression of E- cadherin was negatively correlated with lymphatic metastasis, clinical stage, and cancer differentiation (P < 0.05). Spearman test showed a positive correlation between KiSS- 1 and E- cadherin expression (r(s)=0.722, P < 0.05).
CONCLUSIONKiSS- 1 and E- cadherin may play important roles in inhibiting the invasion and metastasis of gastric cardia carcinoma.
Adult ; Aged ; Aged, 80 and over ; Cadherins ; metabolism ; Cardia ; pathology ; Female ; Humans ; Kisspeptins ; Male ; Middle Aged ; Neoplasm Metastasis ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Suppressor Proteins ; metabolism
3. Comparison of different protocols for protein extraction from formalin-fixed paraffin embedded esophageal squamous cell carcinoma tissues
Ye-Lin JIAO ; Hao-Jie RUAN ; She-Gan GAO ; Yi-Jun QI ; Ye-Lin JIAO ; Yun-Gang ZHAO ; Qi-Wei LIU
Acta Anatomica Sinica 2021;52(1):141-145
Objective To explore protein extraction efficiency from formaldehyde-fixed paraffin embedded (FFPE) esophageal squamous cell carcinoma (ESCC) tissue samples with different protocols. Methods Six different lysis buffers with 100 °C or 105 °C. treatments were used for protein extraction, followed by evaluation of protein quantity and quality with Bradford, sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis, Western blotting and immunohistochemistry (IHC), using 8 FFPE samples of ESCC. Results The optimal method for protein extraction from FFPE ESCC tissue was Laemmli lysis buffer (Buffer 4) treated with 100 °C incubation, evidenced by highest amount of protein recovery. Western blotting and IHC method measured consistent 14-3-3σ expression in FFPE ESCC tissue samples. Protein precipitated by two volumes of acetonitrite acetonitrile(ACN) (0.1% trifluoroacetic acid) relative to protein amount reduced background staining on SDS-PAGE gels by commassie staining. Conclusion Laemmli lysis buffer combined with 100 °C incubation has the highest protein extraction efficiency from FFPE ESCC tissue samples for Western blotting measurement of protein biomarkers, and ACN protein precipitation can further eliminate residual cross- linked protein by FFPE.