Upregulation of FAM134B inhibits endoplasmic reticulum stress‐related degradation protein expression and promotes hepatocellular carcinogenesis

Abstract

Endoplasmic reticulum (ER) stress can stimulate the proliferation and metastasis of hepatocellular carcinoma (HCC) cells while hindering apoptosis and immune system function, but the molecular mechanism of ER stress in HCC has yet to be fully studied. We aim to investigate the molecular mechanism by which FAM134B inhibits autophagy of HCC cells by reducing the expression of ER stress‐related degradation proteins. Clinical samples were collected for this study. Normal liver cell lines HL7702 and Hep3B and Huh7 HCC cell lines were cultured. Construction of FAM134B knockdown cell line. Cell proliferation was measured using the CCK‐8 assay, while cell migration and invasion capabilities were detected using the plate colony formation assay. Flow cytometry was used to detect the apoptosis rate. Transmission electron microscopy was used to observe the formation of autophagosomes. qRT‐PCR and WB detective expression changes related to autophagy proteins. Finally, the expression of the relevant proteins was observed by immunohistochemistry. The expression of FAM134B was significantly increased in human liver cancer tissue and HCC cell lines Hep3B and Huh7. After the lentiviral vector was transfected into Hep3B cells with sh‐FAM134B, results showed that sh‐FAM134B could effectively inhibit Hep3B cell proliferation and promote HCC cell apoptosis. Meanwhile, sh‐FAM134B could effectively induce the autophagy of Hep3B liver cancer cells. Immunohistochemistry results showed that sh‐FAM134B could effectively induce ER stress. FAM134B inhibits HCC cell autophagy and promotes the progression of liver cancer by inhibiting the expression of ER stress‐related degradation factors such as DERL2, EDEM1, SEL1L and HRD1.

The dual effect of endoplasmic reticulum stress in digestive system tumors and intervention of Chinese botanical drug extracts: a review

“…Mao RT et al (2018) reported that ERS elevated the expression of HepG2 chemokines CXCL1, CXCL2, and CXCL3, and decreased the expression of CXCL8 in HCC cells, which may have exacerbated the tumor invasion. Wang H et al (2023) observed that knockdown of FAM134B, a transcriptional response genes to ERS, was able to upregulate the expression of DERL2, EDEM1, SEL1L and HRD1 in HCC cells Hep3B and Huh7, which ultimately reduces apoptosis and promotes tumor growth through the ERS pathway. Moreover, Yan DK et al (2021) found that inhibition of ERS can increase the sensitivity of HCC cells HepG2 and HuH7 to anticancer drug bortezomib and promote apoptosis of cancer cells, which inversely proves that ERS was able to increase drug resistance and survivability in HCC cells.…”

“…Corilagin is a quinone extracted from many botanical drug such as Punica granatum L. with effects such as anticoagulant and antitumor ( Wang X et al, 2023 ). Wu C et al (2021) found that corilagin (10, 20, 40 μM for 24 h) was able to time- and dose-dependently upregulate cleaved Caspase-7 and cleaved Caspase-12 expression, to downregulate GRP78 expression, to inhibit ESC cell ECA-109, KYSE150 and HEEPIC migration, blocking the cell cycle at G0/G1 phase and promoting apoptosis in vitro and ex vivo .…”

“…Isoquercitrin is a flavonoid widely found in many botanical drugs such as Follum Mori, which has pro-healing and anti-tumor effects ( Wang YB et al, 2023 ). Liu J et al (2023) reported that isoquercitrin showed to be effective in the treatment of GC cell lines such as HGC-27, AGS, MKN-45 and SNU-1 by upregulating BAX, cleaved Caspase-3 and Caspase-12, and downregulating BCL-2.…”

“…Quercetin is a flavonoid from botanical drugs such as Styphnolobium japonicum (L.) Schott with anti-inflammatory, antioxidant, and estrogen-like effects ( Wang YT et al, 2023 ). Quercetin (12.5, 25, 50, 100, 200, 400 μmol/L for 48 h) was found to cause a decrease in GRP78, Snail, and Vimentin mRNA and protein expression levels in HCC cells HepG2, inhibiting ERS and its epithelial mesenchymal transition to suppress the proliferation and metastasis of tumor cells ( Li SS et al, 2017 ).…”

“…Osthole, extracted from the fruit of the botanical drug Cnidium monnieri (L.) Cuss, is a coumarin analog possessing anti-inflammatory, antioxidant, antifibrotic, and antitumor effects ( Wang, XW et al, 2023 ; Zhao, LN et al, 2023 ). Pan et al (2019) found that osthole (20, 40, 60, 80, 100 μM for 24, 48, 72 h) concentration-dependently promoted GRP78, CHOP, ATF-4, GADD34, and XBP-1s expression, downregulated Bax expression, blocked cancer cells at G1 phase, inhibited the proliferation of HCC cells SMMC7721 and induced the apoptosis.…”