1.Expression of GNL3 protein in gastric cancer and its effect on cell proliferation,invasion and migration
Xiaolu CHANG ; Siqi ZHANG ; Lianhua ZHU ; Xuelun JIN ; Runzi WANG ; Shuhao WANG ; Junjie PIAO
Chinese Journal of Clinical and Experimental Pathology 2024;40(6):592-597
Purpose To investigate the expression status of GNL3 in gastric cancer,and to explore the role of GNL3 in tumor proliferation,invasion and migration.Methods The ex-pression of GNL3 mRNA in gastric cancer tissues was analyzed by searching database.The expression of GNL3 protein in 51 gastric cancer tissues and 51 adjacent non-tumor tissues was de-tected by immunohistochemistry(IHC)SP method.The correla-tion between GNL3 protein expression and gastric cancer clinical pathological features was analyzed by x2 test.The expression of GNL3 in gastric cancer cells was silenced by transfection of sh-RNA,and the silencing efficiency was verified by Western blot.The effect of silencing GNL3 on the proliferation of gastric cancer cells was examined by CCK-8,colony formation and EdU stai-ning.In addition,wound-healing assay and Transwell assay were performed to detect the effect of GNL3 silencing on cell invasion and migration.Results SangerBox and UALCAN database re-trieval showed that the expression of GNL3 mRNA was signifi-cantly increased in gastric cancer tissues(P<0.01).IHC stai-ning showed that the positive expression rate of GNL3 protein in gastric cancer tissues was 96.1%,and the high expression rate was 78.4%,which was significantly higher than that in adjacent non-tumor tissues(74.5%,51.0%,P<0.01).Moreover,the high expression of GNL3 was significantly correlated with lymph node metastasis in gastric cancer patients(x2=4.933,P=0.026).CCK-8,colony formation and EdU staining showed that GNL3 silencing inhibited the proliferation of gastric cancer cell SGC-7901.The wound-healing and Transwell assay showed that GNL3 silencing inhibited the migration and invasion of gastric cancer cell.Conclusion The GNL3 protein is highly expressed in gastric cancer tissues,and closely related to the proliferation,migration and invasion of gastric cancer cells.
2.Establishment of Specific Chromatogram and Thin-layer Identification Method for Kaixinsan
Wenya GAO ; Min FENG ; Chang GAO ; Haiyu ZHAO ; Yanyan ZHOU ; Hongjie WANG ; Ruipeng YU ; Yipeng ZHAO ; Lili ZHANG ; Yanqing GAO ; Yanhong XIAO ; Cuina CHENG ; Xiaolu WEI ; Nan SI ; Baolin BIAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(13):37-44
ObjectiveTo establish the specific chromatogram and thin layer chromatography(TLC) identification method of Kaixinsan(KXS) samples, in order to clarify the key quality attributes and provide reference for the quality evaluation of KXS. MethodHigh performance liquid chromatography(HPLC) specific chromatogram of KXS was developed with YMC Hydrosphere C18 column(4.6 mm×250 mm, 5 μm), the mobile phase was acetonitrile(A)-0.2% formic acid aqueous solution(B) for gradient elution(0-15 min, 2%-20%A; 15-25 min, 20%-25%A; 25-30 min, 25%-30%A; 30-45 min, 30%-31%A; 45-50 min, 31%-44%A; 50-65 min, 44%-45%A; 65-73 min, 45%-75%A; 73-95 min, 75%-100%A; 95-105 min, 100%A; 105-105.1 min, 100%-2%A; 105.1-120 min, 2%A), the detection wavelength was 320 nm. Ultra high performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) was used to identify the chemical components of KXS with electrospray ionization(ESI), negative ion mode and scanning range of m/z 50-2 000. TLC identification methods for Poria and Ginseng Radix et Rhizoma in KXS were established. ResultThere were 11 common peaks in the specific chromatogram of KXS, attributed to Polygalae Radix, Poria and Acori Tatarinowii Rhizoma. Taking peak 9(α-asarone) as the reference peak, the relative standard deviations of the retention times of 15 batches of KXS samples were<0.2%. A total of 34 compounds were identified by UHPLC-LTQ-Orbitrap MS, including terpenoids, phenylpropanoids, oligosaccharides and ketones. The established TLC had good separation and was rapid, reliable, simple, feasible, suitable for the identification of Poria and Ginseng Radix et Rhizoma in KXS. ConclusionThe specific chromatogram and TLC of KXS are stable and reproducible. The material basis of KXS is basically clarified by MS, which can provide a reference for the development and quality control of KXS.