Objective:

2

To evaluate the anti-hepatocellular carcinoma (HCC) activity of total alkaloids from

Gelsemium elegans

Benth. (TAG)

in vivo

and

in vitro

and to elucidate their potential mechanisms of action through transcriptomic analysis.

Methods:

2

TAG extraction was conducted

and the primary components were quantified using high-performance liquid chromatography (HPLC). The effects of TAG (100

150

and 200 μg/mL) on various tumor cells

including SMMC-7721

HepG2

H22

CAL27

MCF7

HT29

and HCT116

were assessed. Effects of TAG on HCC proliferation and apoptosis were detected by colony formation assays and cell stainings. Caspase-3

Bcl-2

and Bax protein levels were detected by Western blotting.

In vivo

a tumor xenograft model was developed using H22 cells. Totally 40 Kunming mice were randomly assigned to model

cyclophosphamide (20

mg/kg)

TAG low-dose (TAG-L

0.5 mg/kg)

and TAG high-dose (TAG-H

1 mg/kg) groups

with 10 mice in each group. Tumor volume

body weight

and tumor weight were recorded and compared during 14-day treatment. Immune organ index were calculated. Tissue changes were oberseved by hematoxylin and eosin staining and immunohistochemistry. Additionally

transcriptomic and metabolomic analyses

as well as quatitative real-time polymerase chain reaction (RT-qPCR)

were performed to detect mRNA and metabolite expressions.

Results:

2

HPLC successfully identified the components of TAG extraction. Live cell imaging and analysis

along with cell viability assays

demonstrated that TAG inhibited the proliferation of SMMC-7721

HepG2

H22

CAL27

MCF7

HT29

and HCT116 cells. Colony formation assays

Hoechst 33258 staining

Rhodamine 123 staining

and Western blotting revealed that TAG not only inhibited HCC proliferation but also promoted apoptosis (

P

<

0.05).

In vivo

experiments showed that TAG inhibited the growth of solid tumors in HCC in mice (

P

<

0.05). Transcriptomic analysis and RT-qPCR indicated that the inhibition of HCC by TAG was associated with the regulation of the key gene CXCL13.

Conclusion:

2

TAG inhibits HCC both

in vivo

and

in vitro

with its inhibitory effect linked to the regulation of the key gene CXCL13.